Retrospective Study of clinical and hematological aspects associated with dogs naturally infected by Hepatozoon canis in Ludhiana,Punjab,India

Objective:To evaluate clinical and hematological aspects of dogs naturally infected with Hepatozoon canis(H.canis)presented at the Small Animal Clinics of Guru Angad Dev Veterinary and Animal Sciences University,Ludhiana.Methods:Blood films of 34 naturally infected dogs were examined for haematological alterations and parasitaemia.Signalment and clinical signs were recorded from the animals.Clinical histories were filled oul during the consultation.Results:Of the 34 positive dogs by Giemsa stained peripheral blood films.88.23%presented parasilaemia by H.canis only,while 11.77%had the combination of H.canis,Babesia sp.and Ehrlichia sp.Young male dogs less than oue-year-old,of nondescript breed,were the most commonly affected.And 26.47%were presented with anorexia/inappetence as the only clinical symptom.Other clinical symptoms were mild to moderate fever,pale mucosae and lethargy;a few were also showing the signs of vomiting and diarrhoea.Haematological alterations showed mainly normochromic-normocytic anaemia,leukocytosis and neutrophilia.Conclusions:The findings of this study substantiate that H.canis caused clinical and haematological alterations of the varied intensity in dogs,even with low parasilaemia.should he taken into consideration.

Inhibition of <i>Ehrlichia canis</i>and <i>Babesia canis</i>transmission among ticks fed together on dogs vaccinated with Bm86 antigen

GAVAC (Heber Biotec S.A, Havana, Cuba) is a commercially available vaccine developed with the Rhipicephalus (Boophilus) microplus Bm86 recombinant antigen. Bm86 is a “concealed” antigen that is present in the plasmatic membrane of tick gut epithelial cells with unknown function so far. It is well known that after vaccination in the last fifteen years in Cuba, there was a significant decrease of babesiosis (Babesia bovis and Babesia bigemina) and anaplasmosis (Anaplasma marginale) in cattle. A reduced transmission capacity of ticks fed on tick-immune animals and humans has been reported for several tick-borne pathogens. Recent experiments have demonstrated that an anti-tick vaccine may contribute to the control of tick-borne pathogens not only by decreasing the exposure of susceptible hosts to ticks, but also by reducing the vector capacity of ticks. In this study, the potential of Bm86 vaccination to interfere with pathogen transmission among ticks was evaluated by using as experimental model the brown dog tick Rhipicephalus sanguineus and the tickborne Babesia canis and Ehrlichia canis pathogens. Dogs, vaccinated and not vaccinated, were infested with pathogen-infected ticks and noninfected nymphs of R. sanguineus. After feeding, the pathogen transmission to newly molted adults from co-feeding uninfected nymphs was studied by conventional PCR and qPCR. Results suggest that the anti-Bm86 antibodies could be able to block the transmission of B. canis and/or E. canis from infected to non-infected ticks.

Molecular detection of <i>Ehrlichia canis</i>in ticks population collected on dogs in Meshkin-Shahr, Ardebil Province, Iran

Canine monocytic ehrlichiosis is a disease which can cause death in dogs and cats by the Ehrlichia canis and transmitted by hard ticks. The main vector is Rhipicephalus sanguineus which is a common tick in Iran and common ectoparasite on dogs. Few researches has been done to show Ehrlichiosis situation in dog’s ticks. Animal husbandry in Meshkin-Shahr County from Ardebil province (North-West Iran) is the main job of people. Specimens were collected on dogs’ ears, neck, shoulder and toes and they were transferred to the Entomology Laboratory, School of Public Health, Tehran University of Medical Sciences. After DNA extraction, Ehrlichia agent was detected by nested-PCR, 16 s rRNA amplification. Determination of sequence homologies have been done in Gen-Bank. 146 ticks were identified which included 29.44% female and 47.94% male. Rhipicephalus sanguineus were the most prevalent ticks. Ehrlichia spp and Anaplasma spp were found in 43.84% of all the specimens containing Anaplasma ovis, and Ehrlichia sp. and Herlichia canis. Rhipicephalus sanguineus is widely spread and the most common ticks on dogs. As far as we know, this is the first report of E. canis in vector from Iran. Nested PCR showed that hard ticks can contain Anaplasma ovis, Ehrlichia spp and E. canis. These results warrant studying on vector competence of ticks for the Ehrlichiosis agents.

Arginine kinase in Toxocara canis:Exon-intron organization,functional analysis of site-directed mutants and evaluation of putative enzyme inhibitors

Objective: To determine exon/intron organization of the Toxocara canis(T. canis) AK(TCAK) and to test green and black tea and several other chemicals against the activity of recombinant TCAK in the guanidino-specific region by site-directed mutants. Methods: Amplification of genomic DNA fragments containing introns was carried out by PCRs. The open-reading frame(1 200 bp) of TCAK(wild type) was cloned into the BamH 1/SalI site of pM AL-c2X. The maltose-binding protein-TCAK fusion protein was expressed in Escherichia coli TB1 cells. The purity of the expressed enzyme was verified by SDS-PAGE. Mutations were introduced into the guanidino-specific region and other areas of pM AL/TCAK by PCR. Enzyme activity was measured with an NADH-linked assay at 25℃ for the forward reaction(phosphagen synthesis). Results: Arginine kinase in T. canis has a seven-exon/six-intron gene structure. The lengths of the introns ranged from 542 bp to 2 500 bp. All introns begin with gt and end with ag. Furthermore, we measured the enzyme activity of site-directed mutants of the recombinant TCAK. The K_m value of the mutant(Alanine to Serine) decreased indicating a higher affinity for substrate arginine than the wild-type. The K_m value of the mutant(Serine to Glycine) increased to 0.19 mM. The Km value(0.19 mM) of the double mutant(Alanine-Serine to Serine-Glycine) was slightly greater than in the wild-type(0.12 mM). In addition, several other chemicals were tested; including plant extract Azadiracta indica(A. indica), an aminoglycoside antibiotic(aminosidine), a citrus flavonoid glycoside(rutin) and a commercially available catechin mixture against TCAK. Green and black tea(1:10 dilution) produced 15% and 25% inhibition of TCAK, respectively. The extract of A. indica produced 5% inhibition of TCAK. Moreover, green and black tea produced a non-competitive type of inhibition and A. indica produced a mixed-type of inhibition on TCAK. Conclusions: Arginine kinase in T. canis has a seven-exon/six-intron gene structure. However, further studies are needed to identify a specific compound within the extract causing the inhibitory effect and also to determine the molecular mechanisms behind inhibition of arginine kinase in T. canis.

Immunochemical characterization of antigens of Brucella canis and their use in seroprevalence study of canine brucellosis

Objective:To explore immunochemical characterization of antigens of Brucella canis(B. canis),and the use in seroprevalence study of canine brucellosis.Methods:External hot phosphate buffer saline extract(HPBSE) and internal sonicated(SA) antigens were prepared from B.canis strain MEX 51 and imniunochemically characterized.These antigens were used to test 527 serum samples of dogs by 2-mercaptoethanol-tubc agglutination test(2 ME-TAT), agar gel immunodiffusion test(AGID).dot-ELISA and indirect enzyme-linked immunosorbent assay(I-ELISA) to assess the seroprevalence of canine brucellosis.Results:The protein content of HPBSE and SA antigens was 0.387 mg/ml.and 0.195 mg/mL,respectively,whereas carbohydrate content was 0.174 mg/mL and 0.150 mg/mL,respectively.The sodium dodecyl sulfate-polyacrylamide gel electrophoresis(12.5%) of HPBSE and SA,revealed 6 and 8 visible peptide bands ranging from 18-80 kDa and 12-45 kDa,respectively.Western blot analysis showed immunodominant bands of MW 12.28.39 and 45 kl)a for HPBSE and 20-24 kl)a for SA. The AGII) revealed HPBSE as more specific antigen than SA but both I-ELISA and dot-ELISA indicated SA antigen to be more specific and reliable than HPBSE.The seroprevalence of canine brucellosis was 2.27%by 2ME-TAT.1.5%by AGID.3.03%by dot-ELISA and 16.12%by I-ELISA. Conclusions:On the basis of the results of present study,we concluded that HPBSE is suitable antigen for AGID,which is more specific:whereas SA antigen is suitable for I-ELISA,which is highly sensitive.Therefore,initial screening of serum samples should be carried out by I-ELISA followed by confirmation with AGID.

Effect of Milbemycin oxime on Toxocara canis Eggs and Larvae

Toxocara canis(T.canis)is one of the most important zoonotic parasites of dogs.The aim of the present study was to perform an in vitro analysis of the effect of Milbemycin oxime on T.canis eggs following exposure to a concentration gradient of the drug and to determine the inflammatory reaction produced by the infective T.canis larvae in mice.The present study was undertaken using the model nematode,T.canis,to investigate the effect of Milbemycin oxime on T.canis eggs and larvae.T.canis eggs were exposed to a concentration gradient of Milbemycin oxime in vitro,the higher concentration of Milbemycin oxime was,the lower percentage of infective stage larvae was.Light micrographs showed that Milbemycin oxime induced eggs dissolved and eggshell broken.Histological analyses of mice that stained with hematoxylin and eosin(H&E)showed in lower dosing(10-7 and 10-8 g·mL-1)drug-treated groups,atrophy of alveolar space and interalveolar septum thickening appeared,inflammatory infiltrates accompanied with erythrocytes around blood vessels and bronchioles presented.In higher dosing(10-6,10-5 and 10-4 g·mL-1)drug-treated groups,low-grade or no pathological changes occurred,indicating that Milbemycin oxime could obviously decrease the inflammatory reaction produced by the infection of T.canis larvae in vivo.

Factors Determining Coyote (<i>Canis latrans</i>) Diets

Although studies have documented the potential for coyote (Canis latrans) food use to negatively affect wildlife populations and domesticated animals, they are often equivocal, possibly because most are of small spatial extent, and little is known of factors determining coyote diets. Our objectives were to quantify the diet and identify factors determining coyote food use, particularly game species and livestock, over a large spatial and temporal extent. Contents of gastrointestinal tracts were identified from 263 coyotes opportunistically obtained from hunters, trappers, and as road-kills throughout Florida, 2011-2015. We employed logistic regression in an information-theoretic framework to understand determinants of coyote food use. Coyotes were opportunistic and omnivorous foragers with a diverse diet of vegetation, insects, birds, reptiles, amphibians, and more than 25 species of mammals (including important game species and livestock). They commonly consumed 11 food items (Virginia opossum [Didelphis virginiana], non-mast vegetation, feral hog [Sus scrofa], northern raccoon [Procyon lotor], insects, rabbits (Sylvilagus spp.), skunks [Mephitis mephitis and Spilogale putorius], white-tailed deer (Odocoileus virginianus), mast, birds, and cows [Bos taurus]). Food use was determined by coyote age, sex, and body mass, season of the year, deer hunting and fawning seasons, livestock calving season, and coyote collection method and location/region. As coyotes expand their range and numbers, conservationists may find it useful to understand how this opportunistic and adaptable predator uses available food sources to reduce conflict across the landscape.

Antidermatophytic Activity of Essential Oils against Locally Isolated <i>Microsporum canis</i>—Gaza Strip

There is a need for new antimicrobial drugs due to the continuous development of resistance. Natural antimicrobials are of utmost importance due to safety issue and availability. The antifungal activity of four essential oils (Mentha piperta, Thymus vulgaris, Cymbopogon citratus, and Oreganum majoranum oils) against locally isolated Microsporum canis was determined by disc diffusion method and minimum inhibitory concentration was determined using broth dilution method. Mentha piperta oil showed the highest antifungal activity among tested oils in this study. The total inhibition attached when 0.046% was used with M. piperta, the total inhibition obtained with the 0.133% of C. citratus oil, which is accounted as the second essential oils, also T. vulgaris and O. majoranum oils achieved reductions at 0.133%. The present work has revealed that all oils have been used in low concentrations and produced promising results in comparison to the positive control (Clotrimazole).

Effects of Potassium Dichromate on Distribution of Toxocara canis Larvae within Brain of Mice

The frequency of the infective larvae of Toxocara canis, the common parasite of the dog was assessed in brain portions of male Albino mice ingested two doses： 200 mg/Kg and 400 mg/Kg of potassium dichromate-hexavalent （VI） as an indicator. Only 2,000 ova of the parasite were inoculated orally to twenty seven healthy young male mice. Frequency of the parasite was tested 13 weeks post-inoculation. Using bio-statistical analysis （one way analysis of variance） the frequency of detected larvae of both doses in the brain portions of the experimental animals showed an insignificant difference （P〉 0.05） in comparison with control. The results indicate the safety of small and mild intoxication doses of the potassium dichromate on distribution of Toxocara canis in brain portions of mice.

Characterization of insect-fauna of the free-ranging urban dog, Canis domesticus (L.) carcass in tropical region of Pakistan: A tool for forensic entomology

As the authors’ request, the following paper published in Vol.1, No.2, 29-37, 2013 has been withdrawn from the website.

Multifocal granulomata in presumed Toxocara canis infection in adult

Human infection of Toxocara canis in eye is usually an outcome of accidental ingestion of the embryonated eggs. The average age at diagnosis of ocular Toxocariasis is 7.5 years, ranging from 2 to 31 years. It constitutes 1%-2% of uveitis in children. Diagnosis is based upon the clinical features observed in a young patient and confirmed by the presence of specific Ig G in the serum or aqueous humor by Enzyme-linked immunosorbent assay test. We report a case of Presumed Toxocara infection in 45-year-old male which is unique in presentation with multifocal granulomata in retina. Our Pub Med search could not produce case with similar presentation. Probably this is the first reported case of multifocal granulomata in presumed ocular Toxocara in any age

Diagnostic Evaluation of Ehrlichia canis Human Infections

Ehrlichiosis is an infection of the group of Vector Borne Diseases (VBD), caused by different echrlichia species (spp). Ehrlichiae are primarily recognized as causers of animal diseases, later as human diseases—Humane Monocytic (HME) and Humane Granulocytic (HGE) ehrlichiosis. The dogs, cats and rodents have natural reservoirs of ehrlichial agents. Ehcrlichiae are spread to human’s trough, the bites of infected ticks, sometimes resulting from contact with animals, possible to the transfusion. In period from 2008 to 2013, we analyzed 250 patients suspect of VBD. On examination, there were included clinical, laboratory and epidemiological methods. The confirmation of Ehrlichia canis was in serum of 64 patients, thanks to Indirect Immune fluorescent Assay (IIF). The other laboratory and serological methods (Elisa, IIF, and Western blood) are utilized for differential diagnosis, and detection of co-infective forms of diseases. Ehrlichia canis infection has been reported in humans as a cause of serious illnesses characterized by non specific symptoms: fever, headache, musculoskeletal pain, enlarged liver, neurological, hematological dysfunction. Co-infections with E. canis due to causers of different VBD have been documented in humans. In 53 cases, we detected co-infective participation of E. canis with Rickettsia conorii in 24 cases, Coxiella burneti in 6, and Borrelia burgdorferi in 23 cases. The preferred drug for both human ehrlichiosis is doxycycline, except children to 8 years, pregnant females. Ehrlichiosis is a difficult infectious disease to diagnose and prognose because of dominant non specific symptoms and co-infections.

基于红外相机陷阱技术的卡拉麦里山有蹄类自然保护区狼(Canis lupus)的活动节律

通过红外相机陷阱技术,对新疆卡拉麦里有蹄类自然保护区乔木希拜区域野放普氏野马天敌狼的活动节律进行了研究。结果发现,不同月份狼的日活动差异指数α和昼行性指数β均存在极显著差异(α指数:t=8.009,df=7,P<0.01;β指数:t=13.526,df=7,P<0.01),且β=0.45<13/24,表明在研究区狼为夜行性动物。2011年、2012年和2013年季节活动峰型存在显著差异(2011年:χ2=13.087,df=2,P=0.041 1;2012年:χ2=12.452,df=2,P=0.023 4;2013年:χ2=18.998,df=2,P=0.032 6),因此,不同年份不同季节狼的活动节律存在差异。

新疆卡拉麦里山有蹄类自然保护区狼(Canis lupus)与社区牧民冲突的调查分析

近5 a来,卡拉麦里山有蹄类自然保护区狼捕食家畜事件频繁发生,给当地社区牧民造成经济损失并带来心理恐慌,狼与牧民冲突日益尖锐。2012年7—8月,采用入户面对面访谈的方法,调查了卡拉麦里山保护区内狼的危害状况以及当地社区牧民与狼之间的冲突。结果表明:①6个乡的牧民普遍认为在冬牧点或河谷定居点周围有狼,但是其数量有明显的差异;②在卡拉麦里山有蹄类自然保护区狼捕食家畜的顺序是:绵羊、牛、骆驼、山羊和马,狼对绵羊明显存在捕食偏好。每户被狼捕杀家畜的数量与家庭家畜拥有量呈极显著正相关;③狼袭击家畜的时间存在极显著的差异,以夜晚最为频繁,并且多发生在牧民的冬牧场,即在冬季捕食率较高;④狼一般远离道路和人类活动频繁的矿点、旅游点,而在靠近冬牧点和水源点的地方捕食家畜;⑤60.5%的牧民认为狼对草原生态系统是有害的。

新疆准噶尔盆地东南部狼(Canis lupus)的生境选择及其季相变化

采用痕迹样方和对照样方分析卡拉麦里山有蹄类自然保护区狼（Canis lupus）的生境选择及其季相变化。结果表明：春季,狼选择海拔900~1 000 m的山坡和山顶,距离水源2~4 km,道路0.5~1 km,居民点2~4 km及隐蔽级高（0.9~1）的区域;夏季,选择山坡和山沟、海拔900~1 000 m、与水源距离≤2 km、距离道路1.5~3 km、与居民点的距离4~6 km;以及隐蔽级高（0.9~1）的地域;冬季,选择山坡和沙丘、海拔≤800 m、与水源距离≤2 km、与道路的距离1.5~3 km、与居民点的距离2~4 km、隐蔽级在0.5~0.9的区域。主成分分析表明,春季狼生境选择主要受人为干扰的影响,夏季主要受水源因子干扰的影响,冬季主要受食物干扰的影响。

Canine Myofascial Kinetic Lines: A Descriptive Dissection Study Including Related Function and Locomotion and Comparison of the Human and Equine Myofascial Kinetic Lines

Aim: This dissection study was conducted to verify if the Myofascial kinetic lines, outlined in detail in humans and recently documented in horses, were present in dogs. These dynamic lines present rows of interconnected muscles, myofascia and other fascia structures, which influence the biomechanics of the spine and limbs. Methods: Forty-two dogs of different breeds and genders were dissected, imaged, and videoed. Results: Similar kinetic lines were verified in the dog, as described in humans and horses, and additionally, three new branches of the lines were discovered. The kinetic lines described were three superficial lines: Dorsal, Ventral, and Lateral, which all started in the hindlimb and ended in the temporal and occipital regions. These lines act respectively in spinal extension, flexion, and lateral flexion. Three profound lines, which started in the tail and ended in the head. The Deep Dorsal Line followed the transversospinal myofascia. The Deep Ventral Line showed an additional start deep in the medial hind limb, continued in the hypaxial myofascia, and enveloped all the viscera. Also, the Deep Lateral Line started in the hindlimb but parted along the trunk in the deep lateral myofascial structures. Two helical lines crossed the midline two or three times and served to rotate the spine. The Functional Line established a sling from the axilla to the contralateral stifle and presented a new ipsilateral branch. The Spiral Line connected the head and the ipsilateral tarsus and additionally presented a new straight branch. The four front limb lines describe their motion: the Front Limb Protraction and Retraction, Adduction, and Abduction Lines. Conclusion: The canine lines mirrored the equine and human lines with exceptions due to differences in anatomy, foot posture, lumbosacral flexibility, and their biomechanical constitution as predator versus prey animals. Additionally, three new canine branches were verified and described.

Coyote(Canis latrans)in South America:potential routes of colonization

During the last century,the coyote(Canis latrans)has increased its distribution in Central America.Before the 1980s,it had not been recorded in Panama.New records show that coyotes have crossed the Panama Canal,indicating that continues to expand;therefore,there is a possibility that it will reach northern South America.Our objectives were to identify potential coyote colonization routes to South America,and the variables that favor its expansion.We hypothesized that habitat fragmentation benefits coyote expansion.We applied 7 algorithms to model the potential distribution of the coyote,using 196 presence records and 12 variables.The models with better performance were used to generate a consensus model.Using our consensus model and the areas with highest probability of presence,a potential colonization route was generated between Central America and northern South America.This route lies through southern Costa Rica,along the Pacific coast of Panama to the south,to the Andean mountains in northern Colombia.The variables that explained potential coyote distribution were human population density,altitude,and percentage of crops with positive influence,and tropical broadleaf forests with negative in-fluence.These results indicate that human activities and deforestation are related to coyote distribution expansion.Actions can be implemented within the identified route to improve environmental management,in order to avoid the presence of the coyote in the ecosystems of northern South America.

Tinea capitis caused by Microsporum Canis in a child:a case report

We report a tinea capitis caused by Microsporun Canis.A 6-year-old girl who had a history contact with dogs and cats in rural area.The girl suffered from scalp mass,pustules and itching for 2 months.There were two round patches in right scalp which the several pustules scattered on it.The morphology of the colony on the culture medium,the morphological characteristics under the microscope and the results of molecular sequencing confirmed that the girl’s tinea capitis was caused by Microsporun Canis.The girl was cured with the treatment of oral administration of itraconazole and ketoconazole ointment for external use for ten days.

Fecal progesterone and estradiol changes during the breeding season in captive female wolf

Understanding basic reproductive physiology is crucial for the management of both captive and free-ranging wolf. In the present study, we determined hormonal changes during pregnancy and the estrous cycle in captive female wolf by measuring fecal steroids collected during the breeding season with high performance liquid chromatography （HPLC）. These biochemical analyses were validated using chemical devivatization and mass spectrometry, and interpreted along with the behavioral data. All four females undergoing estrus cycles were copulated with their partners and delivered pups successfully. We found that estradiol concen-trations were significantly higher during the estrus cycle than other stages （p0.01） and progesterone was also significantly increased throughout the pregnancy （p0.01）. These hormonal fluctuations demonstrated pregnancy-specific changes in the fecal progesterone and estradiol con-centrations. Patterns of fecal estradiol and progesterone concentrations during estrous cycles were similar to those reported for other canids.

Serological investigation of vector-borne disease in dogs from rural areas of China

Objective:To evaluate the Anaplasma phagocytophilum(A.phagocytophilum),Ehrlichia canis(E.canis,Dirofilaria immitis(D.immitis)(canine heartworm),Borrelia burgdorferi(B.burgdorferi)infections in countryside dogs from Yunnan,Hainan and Anhui provinces.Methods:Serum samples were collected from 26 dogs in Yunnan.Hainan and Anhui provinces.The samples were tested using a commercial ELISA rapid diagnostic assay kit(SNAP^(?)4Dx^(?);IDEXX Laboratories,Inc.U.S.A.).Meaiiwliile,indirect immunofluorescence assay(IFA)recommended by WHO was conducted to delect IgG to A.phagocytophilum.Two methods were analyzed and compared.Results:The number of serologically positive dogs for IgG to A.phagocytophilum was only 2which was from Hainan province and none of the 26 dogs responded positive for E.canu.D.immitis(canine heartworm,and B.burgdorferi by ELISA rapid diagnostic method.The number of serologically positive dogs for IgG to A.phagocytophilum was 13(50%)by IFA method.Data of the two methods were analyzed by statistical software and the difference was statistically significant(P=0.002).Conclusions:It can be concluded that IFA method was more sensitive than ELISA rapid diagnostic method.However,we need conduct further and intensive epidemiology survey on tick-born diseases pathogens including.4.phagocytophilum,E.canis,D.immitis(canine heartworm),and B.burgdorferi which have public health significance.