Search results for: Brucella canis
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 30

Search results for: Brucella canis

30 Proteomic Analysis of Cytoplasmic Antigen from Brucella canis to Characterize Immunogenic Proteins Responded with Naturally Infected Dogs

Authors: J. J. Lee, S. R. Sung, E. J. Yum, S. C. Kim, B. H. Hyun, M. Her, H. S. Lee

Abstract:

Canine brucellosis is a critical problem in dogs leading to reproductive diseases which are mainly caused by Brucella canis. There are, nonetheless, not clear symptoms so that it may go unnoticed in most of the cases. Serodiagnosis for canine brucellosis has not been confirmed. Moreover, it has substantial difficulties due to broad cross-reactivity between the rough cell wall antigens of B. canis and heterospecific antibodies present in normal, uninfected dogs. Thus, this study was conducted to characterize the immunogenic proteins in cytoplasmic antigen (CPAg) of B. canis, which defined the antigenic sensitivity of the humoral antibody responses to B. canis-infected dogs. In analysis of B. canis CPAg, first, we extracted and purified the cytoplasmic proteins from cultured B. canis by hot-saline inactivation, ultrafiltration, sonication, and ultracentrifugation step by step according to the sonicated antigen extract method. For characterization of this antigen, we checked the sort and range of each protein on SDS-PAGE and verified the immunogenic proteins leading to reaction with antisera of B. canis-infected dogs. Selected immunodominant proteins were identified using MALDI-MS/MS. As a result, in an immunoproteomic assay, several polypeptides in CPAg on one or two-dimensional electrophoresis (DE) were specifically reacted to antisera from B. canis-infected dogs but not from non-infected dogs. The polypeptides with approximate 150, 80, 60, 52, 33, 26, 17, 15, 13, 11 kDa on 1-DE were dominantly recognized by antisera from B. canis-infected dogs. In the immunoblot profiles on 2-DE, ten immunodominant proteins in CPAg were detected with antisera of infected dogs between pI 3.5-6.5 at approximate 35 to 10 KDa, without any nonspecific reaction with sera in non-infected dogs. Ten immunodominant proteins identified by MALDI-MS/MS were identified as superoxide dismutase, bacteroferritin, amino acid ABC transporter substrate-binding protein, extracellular solute-binding protein family3, transaldolase, 26kDa periplasmic immunogenic protein, Rhizopine-binding protein, enoyl-CoA hydratase, arginase and type1 glyceraldehyde-3-phosphate dehydrogenase. Most of these proteins were determined by their cytoplasmic or periplasmic localization with metabolism and transporter functions. Consequently, this study discovered and identified the prominent immunogenic proteins in B. canis CPAg, highlighting that those antigenic proteins may accomplish a specific serodiagnosis for canine brucellosis. Furthermore, we will evaluate those immunodominant proteins for applying to the advanced diagnostic methods with high specificity and accuracy.

Keywords: Brucella canis, Canine brucellosis, cytoplasmic antigen, immunogenic proteins

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29 Identification and Characterization of 18S rRNA Gene of Demodex Canis From the Dog Population of Mizoram, India

Authors: Moneesh Thakur, Hridayesh Prasad, Nikitasha Bora, Parimal Roy Choudhary, A. K. Samanta, Sanjeev Kumar

Abstract:

Canine demodicosis is a common parasitic condition which involves dog skin. Demodicosis in dogs is due the prominent growth of Demodex. Out of various canine Demodex spp., Demodex canis is the most often involved species. Canine demodicosis can occur as either a localized or generalized form of demodicosis severely affect the dogs and in non-treated dogs may cause death. This study was planned with the aim to screen and characterize the 18S rRNA gene of isolated Demodex canis. A total of 1200 dogs were screened during this study period. The skin scrapings of all the suspected dogs were examined under a microscope at 100X magnification for the presence of Demodex canis. The skin scrapings positive for Demodex canis were examined using PCR for confirmation. A total of 35 dogs were confirmed a positive result for D. canis based on 18S rRNA gene amplification by PCR. Further, the 18S rRNA gene of isolated Demodex canis was cloned and sequenced for genome analysis. On the sequence analysis, it was found that isolated sequence (GenBank Accession No. MK177513) had close similarity (99.7%) to that of D. canis genotype of China (Accession No. MG372254).

Keywords: PCR, phylogenetic analysis, cloning and sequening, Demodex canis

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28 Differential Antibrucella Activity of Bovine and Murine Macrophages

Authors: Raheela Akhtar, Zafar Iqbal Chaudhary, Yongqun Oliver He, Muhammad Younus, Aftab Ahmad Anjum

Abstract:

Brucella abortus is an intracellular pathogen affecting macrophages. Macrophages release some components such as lysozymes (LZ), reactive oxygen species (ROS) and reactive nitrite intermediates (RNI) which are important tools against intracellular survival of Brucella. The antibrucella activity of bovine and murine macrophages was compared following stimulation with Brucella abortus lipopolysaccharides. Our results revealed that murine macrophages were ten times more potent to produce antibrucella components than bovine macrophages. The differential production of these components explained the differential Brucella killing ability of these species that was measured in terms of intramacrophagic survival of Brucella in murine and bovine macrophages.

Keywords: bovine macrophages, Brucella abortus, cell stimulation, cytokines, Murine macrophages

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27 Detection and Dissemination of Putative Virulence Genes from Brucella Species Isolated from Livestock in Eastern Cape Province of South Africa

Authors: Rudzani Manafe, Ezekiel Green

Abstract:

Brucella, has many different virulence factors that act as a causative agent of brucellosis, depending on the environment and other factors, some factors may play a role more than others during infection and as a result, play a role in becoming a causative agent for pathogenesis. Brucella melitensis and Brucella abortus are considered to be pathogenic to humans. The genetic regularity of nine potential causes of virulence of two Brucella species in Eastern Cape livestock have been examined. A hundred and twenty isolates obtained from Molecular Pathogenesis and Molecular Epidemiology Research Group (MPMERG) were used for this study. All isolates were grown on Brucella agar medium. Nine primer pairs were used for the detection of virB2, virB5, vceC, btpA, btpB, prpA, betB, bpe275, and bspB virulence factors using Polymerase chain reaction (PCR). Approximately 100% was observed for genes BecC and BetB from B. arbotus. While the lowest gene observed was PrpA at 4.6% from B. arbotus. BetB was detected in 34.7%, while virB2 and prpA (0%) were not detected in B. melitensis. The results from this research suggest that most isolates of Brucella have virulence-related genes associated with disease pathogenesis. Finally, our findings showed that Brucella strains in the Eastern Cape Province are extremely virulent as virulence characteristics exist in most strains investigated.

Keywords: putative virulence genes, brucella, polymerase chain reaction, milk

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26 Therapeutic Management of Toxocara canis Induced Hepatitis in Dogs

Authors: Milind D. Meshram

Abstract:

Ascarids are the most frequent worm parasite of dogs and cats. There are two species that commonly infect dogs: Toxocara canis and Toxascaris leonina. Adult roundworms live in the stomach and intestines and can grow to 7 inches (18 cm) long. A female may lay 200,000 eggs in a day. The eggs are protected by a hard shell. They are extremely hardy and can live for months or years in the soil. A dog aged about 6 years, from Satara was referred to Teaching Veterinary Clinical Complex (TVCC) with a complaint of abdominal pain, anorexia, loss of condition and dull body coat with mucous pale membrane. The clinical examination revealed Anaemia, palpation of abdomen revealed enlargement of liver, slimy feel of the intestine loop, diarrhea.

Keywords: therapeutic management, Toxocara canis, induced hepatitis, dogs

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25 Molecular Identification and Genotyping of Human Brucella Strains Isolated in Kuwait

Authors: Abu Salim Mustafa

Abstract:

Brucellosis is a zoonotic disease endemic in Kuwait. Human brucellosis can be caused by several Brucella species with Brucella melitensis causing the most severe and Brucella abortus the least severe disease. Furthermore, relapses are common after successful chemotherapy of patients. The classical biochemical methods of culture and serology for identification of Brucellae provide information about the species and serotypes only. However, to differentiate between relapse and reinfection/epidemiological investigations, the identification of genotypes using molecular methods is essential. In this study, four molecular methods [16S rRNA gene sequencing, real-time PCR, enterobacterial repetitive intergenic consensus (ERIC)-PCR and multilocus variable-number tandem-repeat analysis (MLVA)-16] were evaluated for the identification and typing of 75 strains of Brucella isolated in Kuwait. The 16S rRNA gene sequencing suggested that all the strains were B. melitensis and real-time PCR confirmed their species identity as B. melitensis. The ERIC-PCR band profiles produced a dendrogram of 75 branches suggesting each strain to be of a unique type. The cluster classification, based on ~ 80% similarity, divided all the ERIC genotypes into two clusters, A and B. Cluster A consisted of 9 ERIC genotypes (A1-A9) corresponding to 9 individual strains. Cluster B comprised of 13 ERIC genotypes (B1-B13) with B5 forming the largest cluster of 51 strains. MLVA-16 identified all isolates as B. melitensis and divided them into 71 MLVA-types. The cluster analysis of MLVA-16-types suggested that most of the strains in Kuwait originated from the East Mediterranean Region, a few from the African group and one new genotype closely matched with the West Mediterranean region. In conclusion, this work demonstrates that B. melitensis, the most pathogenic species of Brucella, is prevalent in Kuwait. Furthermore, MLVA-16 is the best molecular method, which can identify the Brucella species and genotypes as well as determine their origin in the global context. Supported by Kuwait University Research Sector grants MI04/15 and SRUL02/13.

Keywords: Brucella, ERIC-PCR, MLVA-16, RT-PCR, 16S rRNA gene sequencing

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24 Epidemiological Survey on Tick-Borne Pathogens with Zoonotic Potential in Dog Populations of Southern Ethiopia

Authors: Hana Tadesse, Marika Grillini, Giulia Simonato, Alessandra Mondin, Giorgia Dotto, Antonio Frangipane Di Regalbono, Bersissa Kumsa, Rudi Cassini, Maria Luisa Menandro

Abstract:

Dogs are known to host several tick-borne pathogens with zoonotic potential; however, scant information is available on the epidemiology of these pathogens in low-income tropical coun- tries and in particular in sub-Saharan Africa. With the aim of investigating a wide range of tick- borne pathogens (i.e., Rickettsia spp., Anaplasma spp., Erhlichia spp., Borrelia spp., Hepatozoon spp. and Babesia spp.), 273 blood samples were collected from dogs in selected districts of Ethiopia and analyzed by real-time and/or end-point PCR. The results of the study showed that Hepatozoon canis was the most prevalent pathogen (53.8%), followed by Anaplasma phagocythophilum (7.0%), Babesia canis rossi (3.3%), Ehrlichia canis (2.6%) and Anaplasma platys (2.2%). Furthermore, five samples tested positive for Borrelia spp., identified as Borrelia afzelii (n = 3) and Borrelia burgdorferi (n = 2), and two samples for Rickettsia spp., identified as Rickettsia conorii (n = 1) and Rickettsia monacensis (n = 1). The finding of Anaplasma phagocythophilum and different species of the genera Borrelia and Rickettsia with zoonotic potential was unexpected and alarming, and calls for further investigation on the roles of dogs and on the tick, species acting as vector in this specific context. Other pathogens (Hepatozoon canis, Babaesia canis rossi, Anaplasma platys, Ehrlichia canis) are already known to have an important impact on the dogs’ health but have minor zoonotic potential as they were rarely or never reported in humans. Dogs from rural areas were found to be at higher risk for different pathogens, probably due to the presence of other wild canids in the same environment. The findings of the present study contribute to a better knowledge of the epidemiology of tick-borne pathogens, which is relevant to human and animal health.

Keywords: Dogs, Tick-borne pathogens, Africa, Ethiopia

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23 Ectoparasites Infestation of Free-Ranging Hedgehog (Etelerix Algirus) in North Western Libya

Authors: M. M. Hosni, A. A. El Maghrbi

Abstract:

The aim of this study was to assess the prevalence of ectoparasites in hedgehogs (Etelerix algirus) in north western region of Libya. Seventy hedgehogs were sampled, and 39 (55.7%) were infested with external parasites. A total of 44 ticks, 491 fleas were collected from the infested hedgehogs and four species of ectoparasites were identified, one mite (Sarcoptes scabiei), one tick (Rhipicephalus appendiculatus) and two fleas (Xenopsylla cheopis and Ctenocephalides canis). For ectoparasites, 10/39 (25.6%) were infested by S. scabiei, 8/39 (20.5%) by Rh. appendiculatus and 11/39 (28.2%) by fleas. The prevalence of mixed infestation with S. scabiei and C. canis was 3(7.7%), Rh. appendiculatus and C. canis was 2 (5.1%) and infestation by two species of fleas was 5 (12.8%). The overall mixed infestation was 10 (25.6%). We concluded that the hedgehogs may play an important role in spreading external parasites and transmission of diseases from one region to another and from wildlife animals to domestic animals and human.

Keywords: ectoparasites, etelerix algirus, hedgehogs, Libya

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22 Isolate-Specific Variations among Clinical Isolates of Brucella Identified by Whole-Genome Sequencing, Bioinformatics and Comparative Genomics

Authors: Abu S. Mustafa, Mohammad W. Khan, Faraz Shaheed Khan, Nazima Habibi

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Brucellosis is a zoonotic disease of worldwide prevalence. There are at least four species and several strains of Brucella that cause human disease. Brucella genomes have very limited variation across strains, which hinder strain identification using classical molecular techniques, including PCR and 16 S rDNA sequencing. The aim of this study was to perform whole genome sequencing of clinical isolates of Brucella and perform bioinformatics and comparative genomics analyses to determine the existence of genetic differences across the isolates of a single Brucella species and strain. The draft sequence data were generated from 15 clinical isolates of Brucella melitensis (biovar 2 strain 63/9) using MiSeq next generation sequencing platform. The generated reads were used for further assembly and analysis. All the analysis was performed using Bioinformatics work station (8 core i7 processor, 8GB RAM with Bio-Linux operating system). FastQC was used to determine the quality of reads and low quality reads were trimmed or eliminated using Fastx_trimmer. Assembly was done by using Velvet and ABySS softwares. The ordering of assembled contigs was performed by Mauve. An online server RAST was employed to annotate the contigs assembly. Annotated genomes were compared using Mauve and ACT tools. The QC score for DNA sequence data, generated by MiSeq, was higher than 30 for 80% of reads with more than 100x coverage, which suggested that data could be utilized for further analysis. However when analyzed by FastQC, quality of four reads was not good enough for creating a complete genome draft so remaining 11 samples were used for further analysis. The comparative genome analyses showed that despite sharing same gene sets, single nucleotide polymorphisms and insertions/deletions existed across different genomes, which provided a variable extent of diversity to these bacteria. In conclusion, the next generation sequencing, bioinformatics, and comparative genome analysis can be utilized to find variations (point mutations, insertions and deletions) across different genomes of Brucella within a single strain. This information could be useful in surveillance and epidemiological studies supported by Kuwait University Research Sector grants MI04/15 and SRUL02/13.

Keywords: brucella, bioinformatics, comparative genomics, whole genome sequencing

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21 Determining the Presence of Brucella abortus Antibodies by the Indirect Elisa Method in Bovine Bulk Milk and Risk Factors in the Peri-Urban Zones of Bamenda Cameroon

Authors: Cha-ah C. N., Awah N. J., Mouiche M. M. M.

Abstract:

Brucellosis is a neglected zoonotic disease of animals and man caused by bacteria of genus Brucella. Though eradicated in some parts of the world, it remains endemic in sub-Saharan Africa including Cameroon. The aim of this study was to contribute to the epidemiology of brucellosis in the North-West region of Cameroon by detecting the presence of anti-Brucella antibodies in bovine bulk milk as this serves as a route of transmission from animals to man. A cross sectional study was conducted to determine the prevalence of Brucella abortus antibodies in bovine bulk milk in the peri-urban zones of Bamenda. One hundred bulk milk samples were collected from 100 herds and tested by milk I-ELISA test. The conducted study revealed the presence of anti-Brucella abortus antibodies in bovine bulk milk. The study revealed that bovine brucellosis is widespread in animal production systems in this area. The animal infection pressure in these systems has remained strong due to movement of livestock in search of pasture, co-existence of animal husbandry, communal sharing of grazing land, concentration of animals around water points, abortions in production systems, locality of production systems and failure to quarantine upon introduction of new animals. The circulation of Brucella abortus antibodies in cattle farms recorded in the study revealed potential public health implication and suggest economic importance of brucellosis to the cattle industry in the Northwest region of Cameroon. The risk for re-emergence and transmission of brucellosis is evident as a result of the co-existence of animal husbandry activities and social-cultural activities that promote brucellosis transmission. Well-designed countrywide, evidence-based studies of brucellosis are needed. These could help to generate reliable frequency and potential impact estimates, to identify Brucella reservoirs, and to propose control strategies of proven efficacy.

Keywords: brucellosis, bulk milk, northwest region Cameroon, prevalence

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20 Current Status and Prospects of Further Control of Brucellosis in Humans and Domestic Ruminants in Bangladesh

Authors: A. K. M. Anisur Rahman

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Brucellosis is an ancient and one of the world's most widespread zoonotic diseases affecting both, public health and animal production. Its current status in humans and domestic ruminants along with probable means to control further in Bangladesh are described. The true exposure prevalence of brucellosis in cattle, goats, and sheep seems to be low: 0.3% in cattle, 1% in goats and 1.2% in sheep. The true prevalence of brucellosis in humans was also reported to be around 2%. In such a low prevalence scenario both in humans and animals, the positive predictive values of the diagnostic tests were very low. The role Brucella species in the abortion of domestic ruminants is less likely. Still now, no Brucella spp. was isolated from animal and human samples. However, Brucella abortus DNA was detected from seropositive humans, cattle, and buffalo; milk of cow, goats, and gayals and semen of an infected bull. Consuming raw milk and unpasteurized milk products by Bangladeshi people are not common. Close contact with animals, artificial insemination using semen from infected bulls, grazing mixed species of animals together in the field and transboundary animal movement are important factors, which should be considered for the further control of this zoonosis in Bangladesh.

Keywords: brucellosis, control, human, zoonosis

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19 Serological Evidence of Brucella spp, Coxiella burnetti, Chlamydophila abortus, and Toxoplasma gondii Infections in Sheep and Goat Herds in the United Arab Emirates

Authors: Nabeeha Hassan Abdel Jalil, Robert Barigye, Hamda Al Alawi, Afra Al Dhaheri, Fatma Graiban Al Muhairi, Maryam Al Khateri, Nouf Al Alalawi, Susan Olet, Khaja Mohteshamuddin, Ahmad Al Aiyan, Mohamed Elfatih Hamad

Abstract:

A serological survey was carried out to determine the seroprevalence of Brucella spp, Coxiella burnetii, Chlamydophila abortus, and Toxoplasma gondii in sheep and goat herds in the UAE. A total of 915 blood samples [n= 222, [sheep]; n= 215, [goats]) were collected from livestock farms in the Emirates of Abu Dhabi, Dubai, Sharjah and Ras Al-Khaimah (RAK). An additional 478 samples (n= 244, [sheep]; n= 234, (goats]) were collected from the Al Ain livestock central market and tested by indirect ELISA for pathogen-specific antibodies with the Brucella antibodies being further corroborated by the Rose-Bengal agglutination test. Seropositivity for the four pathogens is variably documented in sheep and goats from the study area. Respectively, the overall livestock farm prevalence rates for Brucella spp, C. burnetii, C. abortus, and T. gondii were 2.7%, 27.9%, 8.1%, and 16.7% for sheep, and 0.0%, 31.6%, 9.3%, and 5.1% for goats. Additionally, the seroprevalence rates Brucella spp, C. burnetii, C. abortus, and T. gondii in samples from the livestock market were 7.4%, 21.7%, 16.4%, and 7.0% for sheep, and 0.9%, 32.5%, 19.2%, and 11.1% for goats respectively. Overall, sheep had 12.59 more chances than goats of testing seropositive for Brucella spp (OR, 12.59 [95% CI 2.96-53.6]) but less likely to be positive for C. burnetii-antibodies (OR, 0.73 [95% CI 0.54-0.97]). Notably, the differences in the seroprevalence rates of C. abortus and T. gondii in sheep and goats were not statistically significant (p > 0.0500). The present data indicate that all the four study pathogens are present in sheep and goat populations in the UAE where coxiellosis is apparently the most seroprevalent followed by chlamydophilosis, toxoplasmosis, and brucellosis. While sheep from the livestock market were more likely than those from farms to be Brucella-seropositive than those, the overall exposure risk of C. burnetii appears to be greater for goats than sheep. As more animals from the livestock market were more likely to be seropositive to Chlamydophila spp, it is possible that under the UAE animal production conditions, at least, coxiellosis and chlamydophilosis are more likely to increase the culling rate of domesticated small ruminants than toxoplasmosis and brucellosis. While anecdotal reports have previously insinuated that brucellosis may be a significant animal health risk in the UAE, the present data suggest C. burnetii, C. abortus and T. gondii to be more significant pathogens of sheep and goats in the country. Despite this possibility, the extent to which these pathogens may nationally be contributing to reproductive failure in sheep and goat herds is not known and needs to be investigated. Potentially, these agents may also carry a potentially zoonotic risk that needs to be investigated in risk groups like farm workers, and slaughter house personnel. An ongoing study is evaluating the seroprevalence of bovine coxiellosis in the Emirate of Abu Dhabi and the data thereof will further elucidate on the broader epidemiological dynamics of the disease in the national herd.

Keywords: Brucella spp, Chlamydophila abortus, goat, sheep, Toxoplasma gondii, UAE

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18 Analyzing the Visual Capability of the Siberian Husky Breed of the Common Dog (Canis lupus familiaris) to Detect Terminally-Ill Patients Undergoing Palliative Care

Authors: Maximo Cozzetti

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The aim is to evaluate the capability of the 'Siberian Husky' (FCI-Standard Nº 270) breed of the common dog (Canis lupus familiaris) to detect terminally-ill human patients undergoing palliative care. A total of 49 such patients that fulfill the 'National Scientific and Technical Research Council–Ethical Principles for the Behavior of the Scientific and Technical Investigator' policy, (mainly affected with Stage IV Hodgkin lymphoma or Stage IV Carcinoma, though various other terminal diseases were present) and 49 controls were enrolled. A total of 13 specimens of Siberian Huskies (Canis lupus familiaris FCI – Standard Nº 270) were selected. After a conditioning training regime in which the canines were rewarded when identifying terminally ill patients and excluding the control subjects, a double-blind experiment was conducted in which the canines were presented with a previously unknown patient through an olfactory-proof plexiglass window for 2-minute intervals. The test subjects correctly identified 89.80% of the humans as either ‘ill’ or ‘healthy’. It is important to note that both groups of humans were selected considering and preventing confounding and self-identifying factors such as age, ethnicity, clothing, posture, skin color, alopecia (chemotherapy-induced or otherwise), etc. The olfactory-proofing of the test area rules out the use of the sense of smell to detect distinctive drugs or bodily odors that may be associated with terminal diseases. Thus, the Siberian Husky breed of the common dog shows the visual capability to detect and identify terminally ill patients undergoing palliative care regardless of age, posture, and quantity of hair. Though the capability of the breed of dog to detect terminally-ill patients was observed thoroughly during the course of the experiments, the exact process by which the canines identify the test subjects remains unknown and further research is encouraged.

Keywords: Canis lupus familiaris, Siberian Husky, visual identification of terminall illness, FCI-Standard Nº270

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17 Sensitivity and Specificity of Some Serological Tests Used for Diagnosis of Bovine Brucellosis in Egypt on Bacteriological and Molecular Basis

Authors: Hosein I. Hosein, Ragab Azzam, Ahmed M. S. Menshawy, Sherin Rouby, Khaled Hendy, Ayman Mahrous, Hany Hussien

Abstract:

Brucellosis is a highly contagious bacterial zoonotic disease of a worldwide spread and has different names; Infectious or enzootic abortion and Bang's disease in animals; and Mediterranean or Malta fever, Undulant Fever and Rock fever in humans. It is caused by the different species of genus Brucella which is a Gram-negative, aerobic, non-spore forming, facultative intracellular bacterium. Brucella affects a wide range of mammals including bovines, small ruminants, pigs, equines, rodents, marine mammals as well as human resulting in serious economic losses in animal populations. In human, Brucella causes a severe illness representing a great public health problem. The disease was reported in Egypt for the first time in 1939; since then the disease remained endemic at high levels among cattle, buffalo, sheep and goat and is still representing a public health hazard. The annual economic losses due to brucellosis were estimated to be about 60 million Egyptian pounds yearly, but actual estimates are still missing despite almost 30 years of implementation of the Egyptian control programme. Despite being the gold standard, bacterial isolation has been reported to show poor sensitivity for samples with low-level of Brucella and is impractical for regular screening of large populations. Thus, serological tests still remain the corner stone for routine diagnosis of brucellosis, especially in developing countries. In the present study, a total of 1533 cows (256 from Beni-Suef Governorate, 445 from Al-Fayoum Governorate and 832 from Damietta Governorate), were employed for estimation of relative sensitivity, relative specificity, positive predictive value and negative predictive value of buffered acidified plate antigen test (BPAT), rose bengal test (RBT) and complement fixation test (CFT). The overall seroprevalence of brucellosis revealed (19.63%). Relative sensitivity, relative specificity, positive predictive value and negative predictive value of BPAT,RBT and CFT were estimated as, (96.27 %, 96.76 %, 87.65 % and 99.10 %), (93.42 %, 96.27 %, 90.16 % and 98.35%) and (89.30 %, 98.60 %, 94.35 %and 97.24 %) respectively. BPAT showed the highest sensitivity among the three employed serological tests. RBT was less specific than BPAT. CFT showed the least sensitivity 89.30 % among the three employed serological tests but showed the highest specificity. Different tissues specimens of 22 seropositive cows (spleen, retropharyngeal udder, and supra-mammary lymph nodes) were subjected for bacteriological studies for isolation and identification of Brucella organisms. Brucella melitensis biovar 3 could be recovered from 12 (54.55%) cows. Bacteriological examinations failed to classify 10 cases (45.45%) and were culture negative. Bruce-ladder PCR was carried out for molecular identification of the 12 Brucella isolates at the species level. Three fragments of 587 bp, 1071 bp and 1682 bp sizes were amplified indicating Brucella melitensis. The results indicated the importance of using several procedures to overcome the problem of escaping of some infected animals from diagnosis.Bruce-ladder PCR is an important tool for diagnosis and epidemiologic studies, providing relevant information for identification of Brucella spp.

Keywords: brucellosis, relative sensitivity, relative specificity, Bruce-ladder, Egypt

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16 Key Determinants of Human-Wolf (Canis lupus) Conflict in Shabestar County's Villages of East Azerbaijan Province, Iran

Authors: Nader Habibzadeh

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Developing effective and well-targeted conservation strategies is dependent upon fully understanding the complexities of the local situation. We attempted to discern the main likely wolf-human conflict contributing variables in households of Shabestar county’s villages. Data were collected through questions in 53 semi-structured interviews in 36 villages across Shabestar district in summer 2014. The results suggested that people who have reportedly suffered livestock depredation and who have alternative income sources to livestock, are likely to be particularly hostile toward wildlife. With rapid assessment of households using these few key variables we are able to identify likely conflict hotspots and target conflict resolution efforts in those villages. Based on these results, the most important initial strategies for reducing conflict would be reducing the number of livestock killed by wolf, increasing opportunities to generate income only from livestock holdings rather than alternative income sources.

Keywords: human-wildlife conflict, wolf (Canis lupus), Shabestar, Iran

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15 Enhanced Anti-Dermatophytic Effect of Nanoparticles Stimulated by Laser and Cold Plasma Techniques

Authors: Salama A. Ouf, Amera A. El-Adly, Abdelaleam H. Mohamed

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Dermatophytosis is the infection of keratinized tissues such as hair, nail and the stratum corneum of the skin by dermatophytic fungi. Infection is generally cutaneous and restricted to the non-living cornified layers because of the inability of the fungi to penetrate the deeper tissues or organs of immunocompetent hosts. In Saudi Arabia, Onychomycosis is the most frequent infection (40.3%), followed by tinea capitis (21.9%), tinea pedis (16%), tinea cruris (15.1%), and tinea corporis (6.7%). Several azole compounds have been tried to control dermatophytic infection, however, the azole-containing medicines may interfere with the activity of hepatic microsomal enzymes, sex and thyroid hormones, and testosterone biosynthesis. In this research, antibody-conjugated nanoparticles stimulated by cold plasma and laser were evaluated in vitro against some dermatophytes isolated from the common types of tinea. Different types of nanomaterials were tested but silver nanoparticles (AgNPs) were proved to be most effective against the dermatophytes under test. The use of cold plasma coupled with antibody-conjugated nano-particles has severe impact on dermatophytes where the inhibition of growth, spore germination keratinase activity was more than 88% in the case of Trichophyton rubrum, T. violaceum, Microsprum canis and M. gypseum. Complete inhibition of growth for all dermatophytes was brought about by the interaction of conjugated nanoparticles, with cold plasma and laser treatment. The in vivo test with inoculated guinea pigs achieved promising results where the recovery from the infection reached 95% in the case of M. canis –inoculated pigs treated with AgNPs pretreated with cold plasma and laser.

Keywords: cold plasma, dermatophytes, laser, silver nanoparticles

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14 Dermatophytoses: Spectrum Evolution of Dermatophytes in Sfax, Tunisia, Between 1999 and 2019

Authors: Khemakhem Nahed, Hammami Fatma, Trabelsi Houaida, Neji Sourour, Sellami Hayet, Makni Fattouma, Turki Hamida, Ayadi Ali

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Dermatophytoses are considered a public health problem and represent 10% of dermatological consultations in our region. Their epidemiology is influenced by various factors, such as lifestyle, human migration patterns, changes in the environment and the host relationship. The understanding of epidemiology has a major impact on their prevention and treatment. The aim of the study is to determine the prevalence pattern of aetiological agents and to describe the clinical characteristics of dermatophytoses between 1999 and 2019. Out of 65 059 subjects suspected to have superficial mycoses, 36 220 (55.67%) were affected with dermatophytoses. The mean age was 40.1 years (range: 10 days to 99 years). The sex ratio was 0.8. Our patients were from urban regions in 80.9% of cases. The most common type of infection was onychomycosis (42.64%), followed by tinea pedis (20.8%), intertrigo (18.3%), tinea corporis (8.48%) and tinea capitis (7.87%). The most isolated dermatophyte was Trichophyton rubrum (76.5%), followed by T. mentagrophytes complex (6.3%), Microsporum canis (5.8%), T. violaceum (5.3%), T. verrucosum (0.83%) and Epidermophyton floccosum (0.3%). Zoophilic agents have become more prevalent and their frequency has been increased from 6.46% in 1999 to 13% in 2019. It is interesting to note that M. canis has been on the rise since 2010 and it was the first etiological agent of tinea capitis (48%), while infections caused by T. violaceum continued to decrease from 1999 (16.2%) to 2019 (4.7%). Other dermatophytes have been rarely isolated: T. tonsurans (9 cases), T. schoenleinii (3 cases), T. soudanense (2 cases), M. fulvum (1 case), M. audouinii (1 case) and M. ferrugineum (2 cases).T. mentagrophytes var. quinckeanum was isolated from an inflammatory tinea capitis lesion in an a-3-year-old girl. T. mentagrophytes var. erinacei was isolated from the first case of tinea manuum, in-a-10-year-old girl. The same fungus was isolated from the hair and scales of the hedgehog. Our study showed significant changes in the dermatophytes spectrum in our region. The prevalence of zoophilic species increased in recent years due to people's behavioral changes with the adoption of pets and animal husbandry in urban settings. Molecular methods are often crucial that help us to refine the identification strains of dermatophytes and to identify their origin of the contamination.

Keywords: dermatophytoses, PCR-sequencing, spectrum, Sfax, Tunisia

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13 Distribution and Population Status of Canis spp. Threats and Conservation in Lehri Nature Park, Salt Range, District Jhelum

Authors: Muhammad Saad, AzherBaig, Anwar Maqsood, Muhammad Waseem

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The grey wolf has been ranked endangered and Asiatic jackal as near threatened in Pakistan. Scientific data on population and threats to these species are not available in Pakistan, which is required for their proper management and conservation. The present study was conducted to collect data on distribution range, population status and threats to both of these Canis species in Lehri Nature Park. The data were collected using direct observations and indirect signs in the field. The population of grey wolf and Asiatic jackal were scattered into pocket of the study area and its surroundings. The current population of grey wolf was estimated 06 individuals and that of Asiatic jackal 28 individuals in the study area. The present study showed that grey wolf and Asiatic jackal were distributed in the northern and southern part of the study area having dense vegetation cover of tress and shrub between the altitudes of 330 m and 515 m. The research finding revealed that the scrub forest is the most preferred habitat of both the species but due to anthropogenic pressure the scrub forest is under severe threat. The dominant trees species were Acacia modesta, Zizyphus nummularia, and Prosopis juliflora and shrubs species of Dodonea-viscosa, Calotropis procera and Adhatoda vasica. Urial is one of the natural prey species: their population is low due to a number of reasons and therefore the maximum dependence of the wolves was on the livestock of the local and nomadic shepherds. The main prey species in the livestock was goats and sheep. The interviews were conducted with the eye witnesses of wolf attacks including livestock being killed by 5-6 numbers of wolves in different hamlets in the study area. The killing rate of the livestock by the wolves was greater when the nomadic shepherds were present in the area and decreased when they left the area. Presence of nomadic shepherds and killing rate has relation with the shifting of the wolves from the study area. It is further concluded that the population of the grey wolf and Asiatic jackal has decreased over time due to less availability of the natural prey species and habitat destruction.

Keywords: wildlife ecology, population conservation, rehabilitation, conservation

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12 Synthesis, Characterization and Anti-Microbial Study of Urethanized Poly Vinyl Alcohol Metal Complexes

Authors: Maha A. Younus, Dhefaf H. Badri, Maha A. Al Abayaji, Taha M. Salih

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Polymer metal complexes of poly vinyl alcohol and Cu (II), Ni (II), Mn (II) and Co (III) were prepared from the reaction of PVA with three different percentages of urea. The compound was characterized by fourier transform infrared spectrometry (FTIR) analysis and differential scanning calorimetric (DSC) Analysis. It has been established that the polymer and its metal complexes showed good activities against nine pathogenic bacteria (Escherichia coli, Klebsiellapneumonae, Staphylococcusaureus, Staphylococcus Albus, Salmonella Typhoid, Pseudomonas Aeruginosa, Shigella Dysentery, Proteus Morgani, Brucella Militensis). The polymer metal complexes show activity higher than that of the free polymer. The increasing activities were in the order (polymer < pol-Mn< pol-Co < pol-Ni ˂ pol-Cu). The ability of these compounds to show antimicrobial properties suggests that they can be further evaluated for medicinal and/or environmental applications.

Keywords: antimicrobial activity, PVA, polymer-metal complex, urea

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11 Epidemiological, Clinical and Bacteriological Profile of Human Brucellosis in the District of Tunis

Authors: Jihene Bettaieb, Ghassen kharroubi, Rym mallekh, Ines Cherif, Taoufik Atawa, Kaouther Harrabech

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Brucellosis is a major worldwide zoonosis. It is a reportable condition in Tunisia where the disease remains endemic, especially in rural areas. The aim of this study was to describe the epidemiological, clinical, and bacteriological profile of human brucellosis cases notified in the district of Tunis. It was a retrospective descriptive study of cases reported in the district of Tunis through the national surveillance system between the 1st January and 31th December 2017. During the study period, 133 brucellosis confirmed cases were notified. The mean age was 37.5 ± 18.0 years, and 54.9% of cases were males. More than four-fifths (82.7%) of cases were reported in spring and summer with a peak in the month of May (36 cases). Fever and sweats were the most common symptoms; they occurred in 95% and 72% of cases, respectively. Osteoarticular complications occurred in 10 cases, meningitis in one case and endocarditis in one other case. Wright agglutination test and Rose Bengale test were positive in 100% and 91% of cases, respectively. While blood culture was positive in 9 cases and PCR in 2 cases. Brucella melitensis was the only identified specie (9 cases). Almost all cases (99.2%) reported the habit of consuming raw dairy products. Only 5 cases had a suspect contact with animals; among them, 3 persons were livestock breeders. The transmission was essentially due to raw dairy product consumption. It is important to enhance preventive measures to control animal Brucellosis and to educate the population regarding the risk factors of the disease.

Keywords: brucellosis, risk factors, surveillance system, Tunisia

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10 Proteomics Application in Disease Diagnosis and Reproduction İmprovement in Cow

Authors: Abdollah Sobhani, Hossein Vaseghi-Dodaran

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Proteomics is defined as the study of the component of a cell, tissue and biological fluid. This technique has the potential to identify protein biomarkers of a disease states. In this study which was performed on bovine ovarian follicular cysts (BOFC), eight proteins are over expressed in BOFC that these proteins could be useful biomarkers for BOFC. The difference between serum proteome pattern cows affected by postpartum endometritis with healthy cows revealed that concentrations orosomucoid was decreased in endometritis. The comparison proteome of brucella abortus between laboratory adapted strains and clinical isolates could be useful to better understand this disease and vaccine development. Proteomics experiments identified new proteins and pathways that may be important in future hypothesis-driven studies of glucocorticoid-induced immunosuppression. Understanding the molecular mechanisms of effective parameters on male fertility is essential for obtaining high reproductive efficiency by decreasing cost and time. The investigations on proteome of high fertility spermatozoa indicated that expression of some proteins such as casein kinase 2 (CKII) prime poly peptide and tyrosine kinase in high fertility spermatozoa was higher compared to low fertility spermatozoa. Also, some evidence has indicated that variation in protein types and amounts in seminal fluid regulates fertility indexes in dairy bull. In conclusion, proteomics is a useful technique for discovering drugs, vaccine development, and diagnosis disease by biomarkers and improvement of reproduction efficiency.

Keywords: proteomics, reproduction, biomarker, immunity

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9 Integrated Microsystem for Multiplexed Genosensor Detection of Biowarfare Agents

Authors: Samuel B. Dulay, Sandra Julich, Herbert Tomaso, Ciara K. O'Sullivan

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An early, rapid and definite detection for the presence of biowarfare agents, pathogens, viruses and toxins is required in different situations which include civil rescue and security units, homeland security, military operations, public transportation securities such as airports, metro and railway stations due to its harmful effect on the human population. In this work, an electrochemical genosensor array that allows simultaneous detection of different biowarfare agents within an integrated microsystem that provides an easy handling of the technology which combines a microfluidics setup with a multiplexing genosensor array has been developed and optimised for the following targets: Bacillus anthracis, Brucella abortis and melitensis, Bacteriophage lambda, Francisella tularensis, Burkholderia mallei and pseudomallei, Coxiella burnetii, Yersinia pestis, and Bacillus thuringiensis. The electrode array was modified via co-immobilisation of a 1:100 (mol/mol) mixture of a thiolated probe and an oligoethyleneglycol-terminated monopodal thiol. PCR products from these relevant biowarfare agents were detected reproducibly through a sandwich assay format with the target hybridised between a surface immobilised probe into the electrode and a horseradish peroxidase-labelled secondary reporter probe, which provided an enzyme based electrochemical signal. The potential of the designed microsystem for multiplexed genosensor detection and cross-reactivity studies over potential interfering DNA sequences has demonstrated high selectivity using the developed platform producing high-throughput.

Keywords: biowarfare agents, genosensors, multipled detection, microsystem

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8 Seroprevalence of Bovine Brucellosis and its Public Health Significance in Selected Sites of Central High Land of Ethiopia

Authors: Temesgen Kassa Getahun, Gezahegn Mamo, Beksisa Urge

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A cross-sectional study was conducted from December 2019 to May 2020 with the aim of determining the seroprevalence of brucellosis in dairy cows and their owners in the central highland of Oromia, Ethiopia. A total of 352 blood samples from dairy cattle, 149 from animal owners, and 17 from farm workers were collected and initially screened using the Rose Bengal Plate test and confirmed by the Complement Fixation test. Overall seroprevalence was 0.6% (95% CI: 0.0016–0.0209) in bovines and 1.2% (95% CI: 0.0032–0.0427) in humans. Market-based stock replacement (OR=16.55, p=0.002), breeding by artificial insemination (OR=7.58, p=0.05), and parturition pen (OR = 11.511, p=0.027) were found to be significantly associated with the seropositivity for Brucella infection in dairy cattle. Human housing (OR=1.8, p=0.002), contact with an aborted fetus (OR=21.19, p=0.017), drinking raw milk from non-aborted (OR=24.99, p=0.012), aborted (OR=5.72, p=0.019) and retained fetal membrane (OR=4.22, p=0.029) cows had a significant influence on human brucellosis. A structured interview question was administered to 284 respondents. Accordingly, most respondents had no knowledge of brucellosis (93.3%), and in contrast, 90% of them consumed raw milk. In conclusion, the present seroprevalence study revealed that brucellosis was low among dairy cattle and exposed individuals in the study areas. However, since there were no control strategies implemented in the study areas, there is a potential risk of transmission of brucellosis in dairy cattle and the exposed human population in the study areas. Implementation of a test and slaughter strategy with compensation to farmers is recommended, while in the case of human brucellosis, continuous social training and implementing one health approach framework must be applied.

Keywords: abortion, bovine brucellosis, human brucellosis, risk factors, seroprevalence

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7 Retrospective Statistical Study on the Evolution of Brucellosis during the Last Decade (2011-2021) in Medea, Algeria

Authors: Mammar Khames, Mustapha Oumouna

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Brucellosis is one of the most common zoonoses in the world. It represents a serious threat to human health; the existence of brucellosis in Algeria dates back to the beginning of the 19th century. Its transmission to humans is through coccobacilli of the genus Brucella following direct contact with contaminated animals or indirectly through the consumption of their unpasteurized dairy products. The present investigation covers a retrospective study on human brucellosis in the district of Medea over a period from 2011 to 2021 at the level of two public health establishments. In the first place, it is at the level of the Directorate of Public Health and in the infectious department level at Medea Hospital, and at the level of the directorate of agricultural services in the third place. The results showed that during these eleven years of study, 795 cases were collected from the department of health and population, and 141 cases were collected from the infectious department of the district of Medea. A total of 56 cases of bovine brucellosis were obtained from the directorate of agricultural services of the district of Medea. Human brucellosis affects all age groups with different percentages, but the rate has been higher in the 20-44 age group, with a predominance of men. However, the geographic distribution map of the cases shows that the western part of the district was the most affected. A cohort of 141 cases was hospitalized at the infectious service level of Medea Hospital. They were 89 men and 52 women. The most common age reached is [20-44] years. The majority were of rural origin. Two serological reactions were performed for diagnosis: the buffered antigen test and Wright's serodiagnosis. Bovine brucellosis affects all age groups with different percentages, but the rate was higher in the 2-to-4-year age group, with a predominance of females. From these data, we conclude that brucellosis has a strong spread in the region studied.

Keywords: human brucellosis, serology, Medea, Algeria

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6 Livestock Depredation by Large Predators: Patterns, Perceptions and Implications for Conservation and Livelihoods in Karakoram Mountain Ranges

Authors: Muhammad Zafar Khan, Babar Khan, Muhammad Saeed Awan, Farida Begum

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Livestock depredation has greater significance in pastoral societies like Himalaya-Karakoram-Hindu Kush mountain ranges. The dynamics of depredation by large predators (snow leopard and wolf) and its implications for conservation and livelihoods of local people was investigated by household surveys in Hushey valley of Central Karakoram National Park, Pakistan. We found that, during five years (2008-12) 90% of the households in the valley had lost their livestock to snow leopard and wolf, accounting for 4.3% of the total livestock holding per year. On average each household had to bear a loss of 0.8 livestock head per year, equivalent to Pak Rupees 9,853 (US$ 101), or 10% of the average annual cash income. Majority of the predation incidences occurred during late summer in alpine pastures, mostly at night when animals were not penned properly. The prey animals in most of the cases were females or young ones. Of the total predation incidences, 60% were attributed to snow leopard, 37% to wolf, while in 3% the predator was unknown. The fear of snow leopard is greater than that of wolf. As immediate response on predation, majority of the local people (64%, n=99) preferred to report the case to their village conservation committee, 32% had no response while only 1% tended to kill the predator. The perceived causes of predation were: poor guarding practices (77%); reduction in wild prey (13%) and livestock being the favourite food of predators (10%). The most preferred strategies for predator management, according to the respondents were improved and enhanced guarding of livestock (72%), followed by increasing wild prey (18%) and lethal control (10%). To strike a balance between predator populations and pastoral livelihoods, better animal husbandry practices should be promoted including: improved guarding through collective hiring of skilled shepherds; corral improvement and use of guard dogs.

Keywords: Panthera unica, Canis lupus, Karakoram, human-carnivore conflict, predation

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5 Representations of Wolves (Canis lupus) in Feature Films: The Detailed Analysis of the Text and Picture in the Chosen Movies

Authors: Barbara Klimek

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Wolves are one of the most misrepresented species in literature and the media. They’re often portrayed as vicious, man-eating beasts whose main life goal is to hunt and kill people. Many movie directors use wolves as their main characters in different types of films, especially horror, thriller and science fiction movies to create gore and fear. This, in turn, results in people being afraid of wolves and wanting to destroy them. Such cultural creations caused wolves being stalked, abused and killed by people and in many areas they were completely destroyed. This paper analyzes the representations of wolves in the chosen films in the four main portrayed aspects: 1. the overall picture – true versus false, positive versus negative, based on stereotypes or realistic, displaying wolf behavior typical of the species or fake 2. subjectivity – how humans treat and talk about the animals – as subjects or as objects 3. animal welfare – how humans treat wolves and nature, are the human – animal relations positive and appropriate or negative and abusive 4. empathy – are human characters shown to co-feel the suffering with the wolves, do they display signs of empathy towards the animals, do the animals empathize with humans? The detailed analysis of the text and pictures presented in the chosen films concludes that wolves are especially misrepresented in the movies. Their behavior is shown as fake and negative, based on stereotypes and myths, the human – animal relations are shown mainly as negative where people fear the animals and hunt them and wolves stalk, follow, attack and kill humans. It shows that people do not understand the needs of these animals and are unable to show empathy towards them. The article will discuss the above-mentioned study results in detail and will present many examples. Animal representations in cultural creations, including film have a great impact on how people treat particular species of animals. The media shape people’s attitudes, what in turn results in people either respecting and protecting the animals or fearing, disliking and destroying the particular species.

Keywords: film, movies, representations, wolves

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4 Liposomal Antihelmintics in Parasitology

Authors: Nina Ivanova

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More than one third of the population and animals are infected with parasitic helminths. It is especially difficult to cure the larval forms of parasites. The larvae of Hymenolepis nana invade the villi of the intestinal mucosa. Toxocara larvae can live in the liver, heart, lungs, brain, eyes, and pancreas. Commercial antiparasitic drugs cannot guarantee a 100% cure after a single course of treatment, because parasite larvae invade the villi of the intestinal mucosa and the anthelmintics do not reach and kill cestode larvae. The aim of this work was to conduct a study of liposomal antihelminthics on the laboratory animals under the certification received from the Committee on Bioethics and Deontology. It has been checked: 1) anthelmintic activity of the liposomal form of fenasal in experimental hymenolepidosis of white mice (larval stage - Hymenolepis nana). 2) anthelmintic activity of the liposomal form of albendazole in experimental toxocariasis of white mice (in the lungs at the stage of larval migration). Since some helminths cause hemolysis of erythrocytes as we used a mixture of polar lipids developed by us with antihemolytic activity to obtain liposomes Fenasal and albendazole were included in the liposome membrane in the ratio of anthelmintic: lipids 1:10. . The average size of liposomes was 180 nm, and the concentration of lipids in liposomes was 1%. The researches were carried out on white male mice who were infected with Hymenolepis Nana invasional eggs in a peroral way with a doze of 100 eggs per animal. On the 5th day after infection, a liposomal fenasal and commercial fenasal were administered orally for comparison. The animals were observed for 15 days. Before the introduction of liposomes and on the 3rd, 5th, 15th day after the administration of the drug, studies were carried out on the presence of helminths in the organs of animals. The liposomal fenasal, when administered orally, had an anthelmintic effect on Hymenolepis Nana cysticercoids at a dose of 25 mg/kg. The percentage of efficiency was 90.06, 91.36 96.85% on days 3, 5, 15, respectively. For comparison, the commercial activity was at a dose of 200 mg/ml, which is 8 times higher than the dose of liposomal fenasal. To evaluate the anthelmintic effect of the liposomal form of albendazole in experimental toxocariasis of white mice (toxascaris in the lungs at the stage of larval migration), studies were also carried out on white mice. The animals were infected with invasive eggs of Toxocara canis, orally, at a dose of 100 eggs per animal. On the 5th day after infection, the liposomal albendazole was administered orally. The efficacy of the study dosage form was determined by counting Larva mirgans larvae in the lungs. The results obtained showed that the liposomal albendazole had the greatest anthelmintic effect on Toxocara larvae at a dose of 2.0 mg/kg, which was 3.75 times less than the therapeutic one. At the same time, the percentage of efficiency was 93.75% on the 3rd day, and 98.66% on the 5th day.

Keywords: hymenolepis, Toxocara, larvae, liposomes

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3 Analysis of Differentially Expressed Genes in Spontaneously Occurring Canine Melanoma

Authors: Simona Perga, Chiara Beltramo, Floriana Fruscione, Isabella Martini, Federica Cavallo, Federica Riccardo, Paolo Buracco, Selina Iussich, Elisabetta Razzuoli, Katia Varello, Lorella Maniscalco, Elena Bozzetta, Angelo Ferrari, Paola Modesto

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Introduction: Human and canine melanoma have common clinical, histologic characteristics making dogs a good model for comparative oncology. The identification of specific genes and a better understanding of the genetic landscape, signaling pathways, and tumor–microenvironmental interactions involved in the cancer onset and progression is essential for the development of therapeutic strategies against this tumor in both species. In the present study, the differential expression of genes in spontaneously occurring canine melanoma and in paired normal tissue was investigated by targeted RNAseq. Material and Methods: Total RNA was extracted from 17 canine malignant melanoma (CMM) samples and from five paired normal tissues stored in RNA-later. In order to capture the greater genetic variability, gene expression analysis was carried out using two panels (Qiagen): Human Immuno-Oncology (HIO) and Mouse-Immuno-Oncology (MIO) and the miSeq platform (Illumina). These kits allow the detection of the expression profile of 990 genes involved in the immune response against tumors in humans and mice. The data were analyzed through the CLCbio Genomics Workbench (Qiagen) software using the Canis lupus familiaris genome as a reference. Data analysis were carried out both comparing the biologic group (tumoral vs. healthy tissues) and comparing neoplastic tissue vs. paired healthy tissue; a Fold Change greater than two and a p-value less than 0.05 were set as the threshold to select interesting genes. Results and Discussion: Using HIO 63, down-regulated genes were detected; 13 of those were also down-regulated comparing neoplastic sample vs. paired healthy tissue. Eighteen genes were up-regulated, 14 of those were also down-regulated comparing neoplastic sample vs. paired healthy tissue. Using the MIO, 35 down regulated-genes were detected; only four of these were down-regulated, also comparing neoplastic sample vs. paired healthy tissue. Twelve genes were up-regulated in both types of analysis. Considering the two kits, the greatest variation in Fold Change was in up-regulated genes. Dogs displayed a greater genetic homology with humans than mice; moreover, the results have shown that the two kits are able to detect different genes. Most of these genes have specific cellular functions or belong to some enzymatic categories; some have already been described to be correlated to human melanoma and confirm the validity of the dog as a model for the study of molecular aspects of human melanoma.

Keywords: animal model, canine melanoma, gene expression, spontaneous tumors, targeted RNAseq

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2 Neonatology Clinical Routine in Cats and Dogs: Cases, Main Conditions and Mortality

Authors: Maria L. G. Lourenço, Keylla H. N. P. Pereira, Viviane Y. Hibaru, Fabiana F. Souza, João C. P. Ferreira, Simone B. Chiacchio, Luiz H. A. Machado

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The neonatal care of cats and dogs represents a challenge to veterinarians due to the small size of the newborns and their physiological particularities. In addition, many Veterinary Medicine colleges around the world do not include neonatology in the curriculum, which makes it less likely for the veterinarian to have basic knowledge regarding neonatal care and worsens the clinical care these patients receive. Therefore, lack of assistance and negligence have become frequent in the field, which contributes towards the high mortality rates. This study aims at describing cases and the main conditions pertaining to the neonatology clinical routine in cats and dogs, highlighting the importance of specialized care in this field of Veterinary Medicine. The study included 808 neonates admitted to the São Paulo State University (UNESP) Veterinary Hospital, Botucatu, São Paulo, Brazil, between January 2018 and November 2019. Of these, 87.3% (705/808) were dogs and 12.7% (103/808) were cats. Among the neonates admitted, 57.3% (463/808) came from emergency c-sections due to dystocia, 8.7% (71/808) cane from vaginal deliveries with obstetric maneuvers due to dystocia, and 34% (274/808) were admitted for clinical care due to neonatal conditions. Among the neonates that came from emergency c-sections and vaginal deliveries, 47.3% (253/534) was born in respiratory distress due to severe hypoxia or persistent apnea and required resuscitation procedure, such as the Jen Chung acupuncture point (VG26), oxygen therapy with mask, pulmonary expansion with resuscitator, heart massages and administration of emergency medication, such as epinephrine. On the other hand, in the neonatal clinical care, the main conditions and alterations observed in the newborns were omphalophlebitis, toxic milk syndrome, neonatal conjunctivitis, swimmer puppy syndrome, neonatal hemorrhagic syndrome, pneumonia, trauma, low weight at birth, prematurity, congenital malformations (cleft palate, cleft lip, hydrocephaly, anasarca, vascular anomalies in the heart, anal atresia, gastroschisis, omphalocele, among others), neonatal sepsis and other local and systemic bacterial infections, viral infections (feline respiratory complex, parvovirus, canine distemper, canine infectious traqueobronchitis), parasitical infections (Toxocara spp., Ancylostoma spp., Strongyloides spp., Cystoisospora spp., Babesia spp. and Giardia spp.) and fungal infections (dermatophytosis by Microsporum canis). The most common clinical presentation observed was the neonatal triad (hypothermia, hypoglycemia and dehydration), affecting 74.6% (603/808) of the patients. The mortality rate among the neonates was 10.5% (85/808). Being knowledgeable about neonatology is essential for veterinarians to provide adequate care for these patients in the clinical routine. Adding neonatology to college curriculums, improving the dissemination of information on the subject, and providing annual training in neonatology for veterinarians and employees are important to improve immediate care and reduce the mortality rates.

Keywords: neonatal care, puppies, neonatal, conditions

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1 Using Structural Equation Modeling to Measure the Impact of Young Adult-Dog Personality Characteristics on Dog Walking Behaviours during the COVID-19 Pandemic

Authors: Renata Roma, Christine Tardif-Williams

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Engaging in daily walks with a dog (f.e. Canis lupus familiaris) during the COVID-19 pandemic may be linked to feelings of greater social-connectedness and global self-worth, and lower stress after controlling for mental health issues, lack of physical contact with others, and other stressors associated with the current pandemic. Therefore, maintaining a routine of dog walking might mitigate the effects of stressors experienced during the pandemic and promote well-being. However, many dog owners do not walk their dogs for many reasons, which are related to the owner’s and the dog’s personalities. Note that the consistency of certain personality characteristics among dogs demonstrates that it is possible to accurately measure different dimensions of personality in both dogs and their human counterparts. In addition, behavioural ratings (e.g., the dog personality questionnaire - DPQ) are reliable tools to assess the dog’s personality. Clarifying the relevance of personality factors in the context of young adult-dog relationships can shed light on interactional aspects that can potentially foster protective behaviours and promote well-being among young adults during the pandemic. This study examines if and how nine combinations of dog- and young adult-related personality characteristics (e.g., neuroticism-fearfulness) can amplify the influence of personality factors in the context of dog walking during the COVID-19 pandemic. Responses to an online large-scale survey among 440 (389 females; 47 males; 4 nonbinaries, Mage=20.7, SD= 2.13 range=17-25) young adults living with a dog in Canada were analyzed using structural equation modeling (SEM). As extraversion, conscientiousness, and neuroticism, measured through the five-factor model (FFM) inventory, are related to maintaining a routine of physical activities, these dimensions were selected for this analysis. Following an approach successfully adopted in the field of dog-human interactions, the FFM was used as the organizing framework to measure and compare the human’s and the dog’s personality in the context of dog walking. The dog-related personality dimensions activity/excitability, responsiveness to training, and fearful were correlated dimensions captured through DPQ and were added to the analysis. Two questions were used to assess dog walking. The actor-partner interdependence model (APIM) was used to check if the young adult’s responses about the dog were biased; no significant bias was observed. Activity/excitability and responsiveness to training in dogs were greatly associated with dog walking. For young adults, high scores in conscientiousness and extraversion predicted more walks with the dog. Conversely, higher scores in neuroticism predicted less engagement in dog walking. For participants high in conscientiousness, the dog’s responsiveness to training (standardized=0.14, p=0.02) and the dog’s activity/excitability (standardized=0.15, p=0.00) levels moderated dog walking behaviours by promoting more daily walks. These results suggest that some combinations in young adult and dog personality characteristics are associated with greater synergy in the young adult-dog dyad that might amplify the impact of personality factors on young adults’ dog-walking routines. These results can inform programs designed to promote the mental and physical health of young adults during the Covid-19 pandemic by highlighting the impact of synergy and reciprocity in personality characteristics between young adults and dogs.

Keywords: Covid-19 pandemic, dog walking, personality, structural equation modeling, well-being

Procedia PDF Downloads 96