Longitudinal study of an outbreak of Trypanosoma evansi infection in equids and dromedary camels in Israel

An outbreak of trypanosomoasis caused by Trypanosoma evansi involving horses, camels and donkeys occurred in a farm in Israel. A longitudinal study of two outbreak phases was conducted which included clinical monitoring, blood smears, packed cell volume (PCV), serology and polymerase chain reaction (PCR) followed by reverse dot blot (RDB) for the molecular detection of infection. This was the first reported T. evansi outbreak in domestic animals in Israel. Most of the camels on the farm (8/10; 80%) were diagnosed with T. evansi infection whereas infection was less prevalent in the horses (3/7; 43%) and donkeys (6/13; 46%). Clinical disease was evident in 4 camels and 1 horse exhibiting characteristic clinical signs, anemia and parasitemia detected on blood smears and by positive RDB. Six other animals were diagnosed as asymptomatic latent carriers by positive RDB and 6 additional animals were only seropositive and were considered suspected carriers. A significant difference was found in the mean PCV between symptomatic and latent carriers with severe anemia observed only in the symptomatic animals. An anaphylactic-like reaction, fatal in one case, was observed in 2 camels diagnosed with severe trypanosome parasitemia immediately following treatment with melarsenoxide cysteamine. Furthermore, recurrence of infection was documented in one camel 4 months post treatment.     

Immune response and antigen recognition in non-pregnant ewes experimentally infected with Neospora caninum tachyzoites

The cellular and humoral responses as well as the antigen recognition during the acute stage of a Neospora caninum (NC) infection were investigated in non-pregnant ewes. The experimentally infected ewes developed specific lymphoproliferative and humoral responses within 2 weeks post-infection (PI). The magnitude of the cellular response showed large variations between animals. A significant decrease in the proliferative response to Con A mitogen and N. caninum, Toxoplasma gondii (TG) antigens was recorded on day 21 post-infection (PI). The humoral response and the pattern of antigen recognition were similar among infected ewes. Proteins of 44, 42, 40, 39 and 28 kDa were intensively recognized by the infected animals during the experiment. The 42 and 28 kDa antigens should be considered as useful for the diagnostic of N. caninum infection, as the intensity of recognition infection of the other antigens had decreased markedly 8 weeks post-infection. For some antigens a sequential recognition was recorded. The 59, 54 and 38-37 kDa proteins were frequently recognized by infected sera during the first weeks of the infection, but recognition of these antigens was absent or rare at the end of the experiment. These antigens could be related to the acute stage of the infection.     

Experimental infection of weaner sheep with S strain Mycobacterium avium subsp. paratuberculosis

We sought to determine whether infection of recently weaned 12-16-week-old Merino lambs with an Australian S strain M. a. paratuberculosis, at doses consistent with natural exposure, could be detected in the first few months post-inoculation. Such detection would facilitate the use of weaner sheep as sentinel animals for the presence of infectious doses of M. a. paratuberculosis on pastures. In controlled pen trials, oral doses of approximately 10(7)-10(8) viable organisms were demonstrated to be infective, whereas doses below 10(4) organisms failed to produce detectable infection. Mycobacterium avium subsp. paratuberculosis (M. a. paratuberculosis) was isolated from intestinal and/or lymphoid tissues collected at necropsy 7 or 14 weeks after first infection, but there were no associated gross or microscopic lesions. Skin testing with intradermal Johnin detected all three infected lambs at 13 weeks post-infection, and one of the three infected lambs at 6 weeks post-infection, with 100% specificity. Results for whole blood IFN-gamma assay showed some correlation with infection status but lacked specificity. One infected lamb gave a positive result in an ELISA for antibodies to M. a. paratuberculosis, 14 weeks post-infection and 1 week after skin testing. This was the first demonstration of experimental infection with S strain M. a. paratuberculosis in Australian Merino sheep at doses likely to be representative of natural infection. Culture from tissues in the first few months post-exposure could facilitate the use of naive weaner sheep as tracer animals to detect heavy contamination of pastures with M. a. paratuberculosis, but low-level contamination may not be detected in such a system.     

Characterization of the humoral immune response in alpacas (Lama pacos) experimentally infected with Fasciola hepatica against cysteine proteinases Fas1 and Fas2 and histopathological findings

A characterization of the humoral immune response of alpacas to Fasciola hepatica Fas1 and Fas2 antigens, two abundant cysteine proteinases in the excretory/secretory (E/S) products, was performed over the course of 6 months of experimental infection. Six adult alpacas aged 1-2 years old received a single dose of 200 F. hepatica metacercariae; two non-infected alpacas were kept as control group. All infected animals shed eggs 8 weeks post-infection (PI) and the number of flukes recovered at necropsy averaged 41+/-4. The livers of infected animals showed regions with chronic inflammation, granuloma containing parasite eggs, necrosis and cirrhosis. Peripheral eosinophilia in infected animals was greatly enhanced 6 weeks post-infection and later. A single peak of serum glutamic piruvic transaminase (SGPT) was observed 4 weeks PI and serum glutamic oxalacetic transaminase (SGOT) elevated 3 weeks PI and later. Circulating IgG Abs against Fas1 and Fas2 were measured by enzyme-linked immunosorbent assay (ELISA). Fas2-ELISA detected the infection 10 days PI reaching to highest titer on 7-8 weeks PI and kept elevated, until the end of infection. Fas1-ELISA detected the infection 2 weeks PI and followed the same pattern as Fas2-ELISA. Anti Fas2 IgG Abs were in higher titers and showed stronger avidity than anti Fas1 IgG Abs. In addition, rabbit IgG antibodies raised against cysteine proteinase Fas2 showed infiltration of this parasite antigen associated to the degradation of bile ducts and liver parenchyma of infected alpacas. In the present study we have established a F. hepatica experimental infection of alpacas, Fas2 appears to have a role in the pathogenesis of the liver damage in alpacas caused by the liver fluke. Infected alpacas elicited a strong humoral immune response against fluke cysteine proteinases Fas1 and Fas2, which might be considered as candidates for immunodiagnosis and vaccine development against fasciolosis in alpacas.     

Primary experimental infection of riverine buffaloes with Fasciola gigantica.

The clinical course of the primary experimental Fasciola gigantica infection was investigated in riverine buffalo calves of the Murrah breed. Nine male calves aged 12-15 months were randomly assigned to two groups of five (Group I) and four (Group II) animals. Each animal in Group I, was orally infected with 1000 metacercariae (mc) of F. gigantica, whereas Group II animals did not receive any infection dose and served as uninfected controls. No clinical signs of fasciolosis were observed until the sixth week post-infection (PI). Group I animals, however, developed recognised symptoms of acute fasciolosis, comprising apyrexic inappetance, anemia, poor weight gain, diarrhoea and sub-mandibular and facial oedema, respectively, from 5, 6, 8, 16 and 17 weeks PI. The signs were intermittent in nature and of variable duration. The prepatent period was of 92-97 days (mean 95.2 +/- 3.1). One of the five infected animals died on Day 147 PI. At necropsy, 36.8 +/- 11.0% of the infection dose was recovered as adult fluke population. The gross lesions were primarily biliary in nature. Group II, the uninfected controls, throughout the study period of 165 days PI, did not show any symptom and were negative for F. gigantica. The study demonstrated that the onset of adverse effects of F. gigantica on the growth and health of the infected host was mainly noted during late prepatency much before coprological prediction and diagnosis. The significance of preventive therapy against fasciolosis during prepatency has been stressed in endemic areas.     

Aspergillosis in camels affected with a specific respiratory and enteric syndrome

During the summer of 1990 deaths, occurred in racing camels (Camelus dromedarius) associated with a specific disease syndrome. Clinical signs included pyrexia, coughing, lachrymation, oedema of the throat and submandibular region and enlargement of submandibular lymph nodes. In terminal cases nervous signs were present and sometimes there was bloody diarrhoea and vomiting. Of 480 camels at least 70 animals were affected with the disease and about 40 died. Morbidity and mortality was greater in camels recently imported. Consistent necropsy findings were extensive petechial and ecchymotic haemorrhage beneath the epicardium, endocardium and visceral pleura and in the mediastinal lymph nodes, and haemorrhagic oedema of the pharyngeal and laryngeal areas. Haemorrhages occurred more variably in abdominal organs and on the omasal and abomasal mucosa. Bronchopneumonia, omasitis and abomasitis were observed on microscopic examination, together with liver and kidney lesions of presumed toxic origin. Fungal hyphae and, occasionally, the characteristic conidial morphology of Aspergillus fumigatus were seen in sections and direct smears from lesions in the respiratory and alimentary tracts. A fumigatus was cultured from trachea, bronchi, bronchioles, lung tissue, heart blood, omasum, abomasum, ileum and submandibular lymph nodes. Whether the role of Aspergillus in the overall syndrome is primary or secondary has not been established; no other potential aetiological agent has been identified.     

Antibody response of calves to a single infection of Fasciola gigantica determined by an indirect fluorescent antibody technique.

The antibody response of six calves infected with 500 metacercariae of Fasciola gigantica each was monitored throughout 30 weeks of infection using an indirect fluorescent antibody technique (IFA). In vitro excysted F gigantica were employed as the test antigen. All animals showed high antibody titres from two to six weeks post-infection. Thereafter the antibody titres diminished gradually. Although all the experimentally infected animals harboured fluke burdens at autopsy, most gave negative IFA tests from 22 weeks post infection onwards. Specific immunofluorescent staining occurred on the surface glycocalyx of the newly excysted flukes. It is likely that this glycocalyx provides one of the earliest antigenic stimuli for the host's immune reactions to fascioliasis.     

Effect of foodstuff contamination by aflatoxin on the one-humped camel (Camelus dromedarius) in Al Ain, United Arab Emirates

Twenty young female adult one-humped racing camels (Camelus dromedarius) kept in camps scattered outside Al Ain city and aged between 3- and 6-years-old, died after a short clinical illness. Affected camels were dull, inappetant and pyrexic, with submandibular oedema and enlargement of submandibular lymph nodes. Of 100 camels within the camps, 31 showed clinical signs. At necropsy examination, the liver of dead animals appeared yellowish, enlarged, congested and friable. The main hepatic histological findings were centrolobular necrosis, haemorrhages and cellular vacuolation. Aflatoxins were detected in sera, liver, ruminal contents and in feed ingested by affected animals. Sera of symptomatic and recovered camels also showed increased levels of glutamic oxaloacetic transaminases, glutamic pyruvic transaminases, aspartate transaminases, gamma glutamyl transaminases, glucose, urea nitrogen, phosphorus and total iron. Decreased levels of albumin, calcium, cholesterol and triglycerides were also observed. It was probable that aflatoxicosis was responsible for clinical signs and subsequent death of the camels. The need for suitable and appropriate storage conditions of animal feed to prevent fungal growth and aflatoxin contamination is highlighted.     

Renal regulation of the excretion of urea in fasting camels

Experiments were performed with young two-humped camels exposed to 36-hour starvation with free access to water. The renal functions were measured by the standard clearance method. In spite of the administration of 20 micrograms DDAVP, a higher urine flow rate was recorded in the camels subjected to control measurements (feed intake) than in the fasting period (1.45 +/- 0.06 vs. 0.96 +/- 0.06 ml . min-1, P less than less than 0.001). On the second day of fasting the camels had a significantly reduced glomerular filtration rate (GFR 317.5 +/- 23.2 vs. 170.2 +/- 17.4 ml . min-1, P less than 0.001), urea output (700.5 +/- 62.9 vs. 352.2 +/- 64.7 mumol . min-1, P less than 0.005), and fractional excretion of urea (26.9 +/- 2.8 vs. 17.9 +/- 1.7%, P less than 0.01), whereas their tubular resorption. of urea (Reab urea/GFR) increased (6.28 +/- 0.61 vs. 9.12 +/- 0.82 mumol . ml-1, P less than 0.02). No significant difference was found in the concentration of urea in plasma in the fed camels and in fasting camels (8.55 +/- 0.64 vs. 11.18 +/- 1.09 mmol . l-1, N. S.). The creatinine inulin clearance ratio (C creat/Cin) was 0.92 +/- 0.07 when the animals were fed and 1.17 +/- 0.05 when the animals starved (P less than 0.001); this suggests that the clearance of endogenous creatinine is not suitable for GFR measurement in camels under different conditions of nutrition. The kidneys of camels regulate the excretion of urea during short-time fasting mainly through the reduction of glomerular filtration rate and just partly through an increased tubular resorption.     

16alpha-Bromo-epiandrosterone therapy modulates experimental feline immunodeficiency virus viremia: initial enhancement leading to long-term suppression

16alpha-Bromo-epiandrosterone (epiBr), a synthetic derivative of the natural hormone dehyroepiandrosterone (DHEA), was evaluated for its effects on feline immunodeficiency virus (FIV) infection in experimental cats. The rationale for this study was based on the ability of DHEA to significantly reduce the mortality to viral infections in mice. DHEA and epiBr also have demonstrable in vitro anti-viral activity for both HIV-1 and FIV. Preliminary pharmacokinetic studies in cats demonstrated that subcutaneously injected epiBr was rapidly absorbed, completely metabolized, and nontoxic. Metabolites were excreted in both urine and feces, with the latter having the most complex pattern of breakdown products. Cats were then divided into four groups; two groups were infected with FIV and two uninfected. Two groups, one infected and one uninfected were treated on 5 consecutive days of weeks 0, 4, 8, 12 and 16 with epiBr. The remaining two groups were mock treated with the drug vehicle alone. Treatment started 1 week prior to infection and extended for 4 weeks after infection. Cats were observed for 20 weeks post-FIV infection. Infected cats had identical decreases in blood neutrophil and lymphocyte counts following, regardless of whether they were treated with epiBr or vehicle alone. The CD4/CD8 T-cell ratio was decreased following FIV exposure, but was significantly more decreased for the epiBr treated animals from week 2 post-infection onward. CD4+ T cells were decreased in FIV-infected cats treated with epiBr compared to their untreated cohort, while CD8+ T cells tended to be higher in treated animals. FIV infected cats that were treated with epiBr had over one-log higher virus loads at week 2 post-infection than non-epiBr treated cohorts. In spite of this enhanced initial viremia, the subsequent levels of virus in the blood were significantly lower in epiBr treated versus untreated animals. EpiBr treated cats had significantly higher FIV-p24 antibody responses than control cats receiving vehicle alone, although primary and secondary antibody responses to a T-cell dependent non-FIV antigen, keyhole limpet hemocyanin (KLH), were unaffected. EpiBr treatment significantly decreased the expected FIV-induced suppression of IL-12 p40 mRNA levels in peripheral blood mononuclear cells (PBMCs) observed at weeks 4, 5, 8, 9 and 16 post-infection, but had no influence on FIV-induced changes in IL-4, IL-6, IL-10, IFN-gamma, MIP-1alpha and RANTES.     

Studies on the age resistance of swine to group A rotavirus infection.

To determine whether swine become naturally age resistant to group A rotavirus infection, colostrum-deprived, rotavirus-naive newborn pigs that were raised in isolation (n = 34) were studied. Neonatal pigs and pigs 1, 2, 4, 6, 8, 10, and 12 weeks of age were inoculated orally with group A porcine rotavirus or mock inoculum and euthanatized at 24, 31, or 48 hours post-infection. Nine sections of small intestine, cecum, and colon were harvested and immunohistochemically examined for evidence of rotavirus replication within enterocytes. Infectivity was semiquantified by intestinal segment, and a composite score was obtained for each animal. In pigs inoculated at 1 week of age, enterocyte infection was mild and scattered; all other pigs became infected regardless of age or region of intestine, and older animals that became infected had infectivity scores similar to those of younger animals. In a second more limited study, pigs raised in the same isolation environment (n = 11) but previously exposed to virus and demonstrating rotavirus serum antibody had a much lower degree of enterocyte infection at 8, 10, and 12 weeks of age (2, 4, and 6 weeks, respectively, after initial exposure to virus). Age resistance to clinical rotavirus disease in swine is due to factors other than an age-dependent development of resistance of enterocytes to infection, at least through 12 weeks of age.     

Vaccination with an autogenous bacterin fails to prevent colonization by Brachyspira intermedia in experimentally infected laying chickens

Avian intestinal spirochaetosis (AIS) is a disease complex affecting adult laying and breeding chickens associated with infection by anaerobic intestinal spirochaetes of the genus Brachyspira. Options for control of AIS are limited, as few effective antimicrobial agents are registered for use in laying chickens. One of the two most commonly encountered pathogenic species in AIS is B. intermedia, and the aim of the current study was to determine whether a B. intermedia bacterin vaccine would help control AIS caused by this species. An autogenous bacterin was prepared from B. intermedia strain HB60 and given twice intramuscularly at a 3-week interval to 12 laying chickens housed in individual cages. Twelve non-vaccinated control chickens were placed in adjacent cages in the same room. Two weeks after the second vaccination all the chickens were experimentally challenged with B. intermedia HB60 by crop tube. Subsequently faeces were cultured for spirochaetes every 2-3 days, faecal water content and chicken weight were measured weekly, and egg numbers and weights were recorded daily. Serum was taken prior to both vaccinations, at the time of challenge and at euthanasia. The chickens were killed 6 weeks post-challenge. The vaccinated chickens showed seroconversion to the vaccine, but antibody levels declined significantly post-infection. In comparison, the non-vaccinated chickens showed seroconversion post-infection. The reason for the reduction in the antibody levels in the vaccinated chickens after infection was not explained. At some point all the chickens excreted spirochaetes in their faeces, and the duration of excretion was not different between vaccinated and non-vaccinated chickens. There were no differences in faecal water content, chicken weights, egg production, or gross and microscopic caecal lesions between vaccinated and non-vaccinated chickens. In conclusion, an autogenous bacterin vaccine did not prevent infection with B. intermedia in laying chickens.     

Study on the susceptibility of Sahel goats to experimental Trypanosoma vivax infection

Sahel goats, also known as Borno whites are found in the northern semi-arid, tsetse free Sahel region of Nigeria. They are transported alongside cattle from this zone to all other zones in the country, including the tsetse-infested zones, for commercial purposes and are kept for some time in these tsetse-infested zones until they are sold. This study therefore assessed the susceptibility of this breed of goats to trypanosome infection and its response to treatment with Berenil. Six bucks were inoculated intravenously with Trypanosoma vivax through the jugular vein while two served as uninfected control. The mean pre-patent period was 4.5 days and increasing parasitaemia followed the establishment of infection. Onset of parasitaemia was associated with increase in rectal temperature in all the infected goats and the temperature peak coincided with the only parasitaemic peak second week post-infection. The infected goats were treated with Berenil (Hoechst, Germany) 3.5mg/kg body weight at 4 weeks post-infection. The packed cell volume (PCV) continued to fall from a mean 30.73+/-0.01% pre-infection to a mean 13.21+/-0.18% at 1 week post-treatment. Deaths were recorded for 4 of the infected goats 1 week post-treatment while the remaining two died 2 weeks post-treatment, not responding to treatment.     

Humoral and cellular immune responses to Fasciola gigantica experimental infection in buffaloes

Humoral and cellular immune responses to Fasciola gigantica experimental infection in buffaloes were studied. The results showed that 33.4+/-9.1% of the infection dose was recovered as adult flukes from infected animals at necropsy. Significant differences of weight gain between infected and non-infected buffaloes was observed at 4 MPI (months post-infection). Anti F. gigantica excretory-secretory products (FgESP)-IgG levels increased significantly from 3 WPI (weeks post-infection) and displayed a peak at 13 WPI. Western blot indicated that in FgESP six major bands of 11.5, 19.0, 23.4, 29.8, 47.5 and 53.2kDa were recognized by F. gigantica-infected buffaloes sera after 0 WPI. Eosinophil numbers increased significantly from 3 WPI in F. gigantica-infected buffaloes and displayed a peak at 8 WPI. Peripheral blood mononuclear cells (PBMC) proliferation induced by FgESP increased from 2 WPI with a peak at 5 WPI. IFNgamma secretion by FgESP-stimulated PBMC appeared early from 1 WPI with three peaks at 2, 5 and 8 WPI, respectively. IL-10 production was observed from 2 WPI with two peaks at 4 and 9 WPI, respectively. Our results suggested that buffaloes were highly susceptible to F. gigantica infection, and this susceptibility could be associated with the late and weak cellular immune response in the early phase of infection and the Th0-like response throughout the infection.     

Functions of neutrophils in sheep experimentally infected with Ehrlichia phagocytophila

Ehrlichia phagocytophila infection in sheep is characterized by persistent neutropaenia, indicative of decreased phagocytic capacity. This predisposes infected animals to other infections. A whole blood flow cytometrical method was used to document the degree and extent of reduced phagocytic and respiratory burst activity in phagocytes during an experimental infection with E. phagocytophila, and monitored until 56 days post-infection. Six sheep at 5 months of age were inoculated with an intravenous injection of infected blood. Six age-matched sheep were used as controls. A period of reduced respiratory burst lasting up to Day 17 post-infection was recorded. The population of cells showing phagocytic activity without respiratory burst was larger in the infected animals compared to controls up to Day 45 post-infection.     

Experimental infection of camels with bluetongue virus

Three camels aged 4–5 years were experimentally infected with Bluetongue virus serotype 1 (BTV-1) and were observed for 75 days. No clinical signs of disease were observed throughout the experiment, however all three animals seroconverted and developed BTV-1 specific neutralising antibodies after challenge. All three camels developed a viraemia from 7 days post infection albeit at a lower level than that usually observed in experimental infections of sheep and cattle. Virus was isolated from the blood of all three animals suggesting that camels may act as a reservoir for BTV and play an important role in its transmission.     

Prophylactic efficacy of buparvaquone in experimentally induced Theileria annulata infection in calves

The antitheilerial activity of buparvaquone (BW 720C) was evaluated in experimentally induced Theileria annulata infections in cross-bred male calves. T. annulata infections were induced by injecting a suspension of infected ground tick tissue suspension (GUTTS) equivalent to two ticks subcutaneously into each calf. Buparvaquone at a dose of 2.5 mg kg-1 body weight was given as a single injection (intramuscularly) on Day 0 (Group 1), Day 8 (Group 2) and Day 12 (Group 3) post-infection. The animals in Groups 4 and 5 were untreated and challenged controls, respectively. All of the recovered animals from Groups 1-4 were challenged with a lethal dose of T. annulata at 6 weeks post-infection. The immunized animals were resistant to the homologous challenge, which killed three of four control animals (Group 5); the controls showed typical antemortem and post-mortem lesions of theileriosis.     

Experimental infections of baboons (Papio spp.) and vervet monkeys (Cercopithecus aethiops) with Trichinella zimbabwensis and successful treatment with ivermectin

Experimental Trichinella zimbabwensis infections were established in three baboons (Papio sp.) and four vervet monkeys (Cercopithecus aethiops) and the clinical-pathological manifestations assessed. The infected animals showed clinical signs ranging from fever, diarrhoea, periorbital oedema and muscular pain in varying degrees. One baboon became blind due to the infection. Levels of creatinine phosphokinase and lactate dehydrogenase increased to reach a peak on Day 42 post-infection (pi) for both baboons and monkeys. Blood parameters such as packed cell volume, levels of red blood cells and white blood cells did not change significantly from the normal ranges except for the levels of eosinophils which peaked above the normal ranges at Day 28 and 56 pi in baboons and at Day 56 pi in monkeys. Two baboons and two monkeys died during the course of the experiment. They were emaciated and showed lesions such as ascites, hydropericardium, congested liver and enlarged gall bladder. Histopathological findings of various muscles included a basophilic transformation of muscle cells, the disappearance of sarcomere myofibrils and basophilic sarcoplasm with the presence of Trichinella larvae in the sarcoplasm. These changes were mainly in the massetter and were of various intensities in the tail, gastrocnemius and biceps muscles. Five consecutive treatments with an oxfendazole-levamisole combination on surviving animals failed to clear the infection whereas ivermectin cleared the infection after one treatment in two monkeys and after two treatments in a baboon.     

Direct and sensitive detection of Trypanosoma evansi by polymerase chain reaction.

The mechanically transmitted haemoflagellate, Trypanosoma evansi causes 'surra', a wasting disease of domestic animals and is highly endemic in distribution in Southeast Asia. The detection of T. evansi is important for improving the epizootiological and animal health status of the region. The specificity and sensitivity of polymerase chain reaction (PCR) using oligonucleotide primers constructed from T. evansi repetitive DNA sequences were studied in the present investigation. Using the assay, it was possible to amplify template DNA of T. evansi derived from buffaloes, camels and horses to a threshold sensitivity level of 0.5 pg and to detect DNA from as few as five organisms in 10 microliters crude blood samples. Following experimental infection of calves with 5 x 10(5) T. evansi, positive signals could be observed as early as 12 h post-infection. DNAs from two common haemoflagellates of cattle, Babesia bigemina and Theileria annulata were not amplified with the primers.     

Zoonotic cases of camelpox infection in India

This study reports the first conclusive evidence of zoonotic camelpox virus (CMLV) infection in humans associated with outbreaks in dromedarian camels (Camelus dromedaries) in northwest region of India during 2009. CMLV infection is usually restricted to camels and causes localised skin lesions but occasionally leads to generalised form of disease. However, the present outbreak involved camel handlers and attendants with clinical manifestations such as papules, vesicles, ulceration and finally scabs over fingers and hands. In camels, the pock-like lesions were distributed over the hairless parts of the body. On the basis of clinical and epidemiological features coupled with serological tests and molecular characterization of the causative agent, CMLV zoonosis was confirmed in three human cases. Clinical samples such as skin scabs/swabs and blood collected from affected animals and humans were analysed initially, for the presence of CMLV-specific antigen and antibodies by counter immunoelectrophoresis (CIE); serum neutralization test (SNT); plaque-reduction neutralization test (PRNT) and indirect immunoperoxidase test which was later confirmed by amplification of CMLV-specific ankyrin repeat protein (C18L) gene. Virus isolation was successful only from samples collected from camels. Further, sequence analyses based on three full-length envelope protein genes (A27L, H3L and D8L) revealed 95.2-99.8% and 93.1-99.3% homology with other Orthopoxviruses at nucleotide and amino acid levels, respectively. Phylogram of the three genes revealed a close relationship of CMLV with Variola virus (VARV). Considering the emerging and re-emerging nature of the virus, its genetic relatedness to VARV, zoonotic potential and productivity losses in camels; the control measures are imperative in curtailing economic and public health impact of the disease. This is the first instance of laboratory confirmed camelpox zoonosis in India.