Hyperprolactinemic African elephant (Loxodonta africana) females exhibit elevated dopamine, oxytocin and serotonin concentrations compared to normal cycling and noncycling, low prolactin elephants†.

Many zoo elephants do not cycle normally, and for African elephants, it is often associated with hyperprolactinemia. Dopamine agonists successfully treat hyperprolactinemia-induced ovarian dysfunction in women, but not elephants. The objective of this study was to determine how longitudinal dopamine, serotonin, and oxytocin patterns in African elephants are related to ovarian cycle function. We hypothesized that dopamine concentrations are decreased, while oxytocin and serotonin are increased in non-cycling, hyperprolactinemic African elephants. Weekly urine and serum samples were collected for eight consecutive months from 28 female African elephants. Females were categorized as follows: (1) non-cycling with average prolactin concentrations of 15 ng/ml or greater (HIGH; n = 7); (2) non-cycling with average prolactin concentrations below 15 ng/ml (LOW; n = 13); and (3) cycling with normal progestagen and prolactin patterns (CYCLING; n = 8). Both oxytocin and serotonin were elevated in hyperprolactinemic elephants. Thus, we propose that stimulatory factors may play a role in the observed hyperprolactinemia in this species. Interestingly, rather than being reduced as hypothesized, urinary dopamine was elevated in hyperprolactinemic elephants compared to CYCLING and LOW prolactin groups. Despite its apparent lack of regulatory control over prolactin, this new evidence suggests that dopamine synthesis and secretion are not impaired in these elephants, and perhaps are augmented.

NEW APPROACH TO STRESS RESEARCH IN PHOCIDS-POTENTIAL OF DEHYDROEPIANDROSTERONE AND CORTISOL/DEHYDROEPIANDROSTERONE RATIO AS MARKERS FOR STRESS IN HARBOR SEALS ( PHOCA VITULINA) AND GRAY SEALS ( HALICHOERUS GRYPUS).

Cortisol is known to reflect the level of the stress response. However, measuring the cortisol concentration only once fails to provide sufficient information about the duration of the stress exposure. Moreover, handling is an acute stressor and increases cortisol secretion especially in wildlife species. Yet, in phocids reliable indicators are missing that reflect potential chronic effects of stress. The adrenal-derived steroid hormone dehydroepiandrosterone (DHEA) has been described as biomarker for the assessment of the stress status but has not been applied to marine mammal stress research. Therefore, DHEA, the sulfated precursor DHEAS and the cortisol/DHEA ratio were determined in serum of different seals. One group consisted of harbor ( Phoca vitulina) and gray seals ( Halichoerus grypus) that were habituated to human handling. The other two groups included healthy and free-ranging seals suffering from a disease. Blood samples were taken from 11 habituated (six males, five females), 17 wild_healthy (13 males, four females), and nine wild_diseased seals (four males, five females). No differences in serum cortisol levels could be detected between wild_healthy and wild_diseased seals. On the contrary, wild_diseased seals showed the lowest DHEA concentration compared to the other two groups. Wild_diseased seals also revealed the highest cortisol/DHEA ratio compared to the habituated_zoo (2,074.7 ± 351.4 vs. 121.5 ± 26.8, P < 0.001) and wild_healthy seals (827.9 ± 214.3, P < 0.01). DHEA and the cortisol/DHEA ratio may reflect differences in the functionality of the hypothalamic-pituitary-adrenal axis and therefore represent valuable tools for the assessment of stress-related effects in seals.

Serological survey of viral diseases relating to reproductive failure among Artiodactyla in Ethiopian Camelus dromedarius.

Serological surveys were performed on Ethiopian camels with a history of abortion to investigate the presence of antibodies against viruses that infect animals classified in the order Artiodactyla. In 2013, 120 serum samples were collected from camels in various parts of Ethiopia. Several viruses related to abortion in ruminants were prevalent. In particular, antibodies against bluetongue virus, were detected at a high rate (76.7% of samples). Additionally, antibodies against Akabane virus and Japanese encephalitis virus were also detected in samples from more than 40% of the camels; however, their antibody titers were relatively low.

Farm animal serum proteomics and impact on human health.

Due to the incompleteness of animal genome sequencing, the analysis and characterization of serum proteomes of most farm animals are still in their infancy, compared to the already well-documented human serum proteome. This review focuses on the implications of the farm animal serum proteomics in order to identify novel biomarkers for animal welfare, early diagnosis, prognosis and monitoring of infectious disease treatment, and develop new vaccines, aiming at determining the reciprocal benefits for humans and animals.

Bartonella spp. in Bats, Guatemala.

To better understand the role of bats as reservoirs of Bartonella spp., we estimated Bartonella spp. prevalence and genetic diversity in bats in Guatemala during 2009. We found prevalence of 33% and identified 21 genetic variants of 13 phylogroups. Vampire bat-associated Bartonella spp. may cause undiagnosed illnesses in humans.

Reference intervals and relationships between health status, carapace length, body mass, and water temperature and concentrations of plasma total protein and protein electrophoretogram fractions in Atlantic loggerhead sea turtles and green turtles.

OBJECTIVE: To determine reference intervals for concentrations of plasma total protein (TP) and electrophoretogram fractions (ELFs) for healthy, wild loggerhead sea turtles (Caretta caretta) and green turtles (Chelonia mydas) and to assess relationships between TP and ELF concentrations and health status, body size, body mass, and water temperature. DESIGN: Evaluation study. ANIMALS: 437 healthy and 35 ill Atlantic loggerhead sea turtles and 152 healthy and 3 ill Atlantic green turtles. PROCEDURES: Free-ranging turtles were captured from a nuclear power plant intake canal in southern Florida. Plasma samples were obtained from all turtles. Plasma TP and ELF concentrations were measured, and reference intervals were calculated. Wilcoxon rank-sum tests were used to compare TP and ELF values between healthy and ill loggerhead sea turtles. Spearman rank correlations were evaluated between concentrations of TP and ELFs and carapace length, body mass, and water temperature. RESULTS: Reference intervals for TP concentrations were 2.2 to 5.2 g/dL and 2.0 to 5.4 g/dL for loggerhead sea turtles and green turtles, respectively. Except for gamma-globulin, concentrations of ELFs were significantly higher in healthy than in ill loggerhead sea turtles. There was a positive correlation between TP, alpha-globulin, beta-globulin, and gamma-globulin concentrations and water temperature in loggerhead sea turtles and between only TP and alpha-globulin concentrations and water temperature in green turtles. CONCLUSIONS AND CLINICAL RELEVANCE: Reference intervals for concentrations of TP and ELFs for healthy, free-ranging loggerhead sea turtles and green turtles can be used in combination with other diagnostic tools to assess health status of sea turtles.

Subolesin vaccination inhibits blood feeding and reproduction of Haemaphysalis longicornis in rabbits.

BACKGROUND: Ticks can transmit numerous tick-borne pathogens and cause a huge economic loss to the livestock industry. Tick vaccines can contribute to the prevention of tick-borne diseases by inhibiting tick infestation or reproduction. Subolesin is an antigenic molecule proven to be a potential tick vaccine against different tick species and even some tick-borne pathogens. However, its effectivity has not been verified in Haemaphysalis longicornis, which is a widely distributed tick species, especially in East Asian countries. Therefore, the purpose of this study was to evaluate the effectivity of subolesin vaccination against H. longicornis in a rabbit model. METHODS: Haemaphysalis longicornis (Okayama strain, female, adult, parthenogenetic strain) and Japanese white rabbits were used as the model tick and animal, respectively. The whole open reading frame of H. longicornis subolesin (HlSu) was identified and expressed as a recombinant protein using E. coli. The expression was verified using sodium dodecyl sulfate polyacrylamide gel electrophoresis, and the immunogenicity of rHlSu against anti-H. longicornis rabbit serum was confirmed using Western blotting. After vaccination of rHlSu in rabbits, experimental infestation of H. longicornis was performed. Variables related to blood-feeding periods, pre-oviposition periods, body weight at engorgement, egg mass, egg mass to body weight ratio, and egg-hatching periods were measured to evaluate the effectiveness of subolesin vaccination. RESULTS: The whole open reading frame of HlSu was 540 bp, and it was expressed as a recombinant protein. Vaccination with rHlSu stimulated an immune response in rabbits. In the rHlSu-vaccinated group, body weight at engorgement, egg mass, and egg mass to body weight ratio were statistically significantly lower than those in the control group. Besides, egg-hatching periods were extended significantly. Blood-feeding periods and pre-oviposition periods were not different between the two groups. In total, the calculated vaccine efficacy was 37.4%. CONCLUSIONS: Vaccination of rabbits with rHlSu significantly affected the blood-feeding and reproduction in H. longicornis. Combined with findings from previous studies, our findings suggest subolesin has the potential to be used as a universal tick vaccine.

[Design and implementation of a national serum bank for the surveillance of exotic animal diseases in the Republic of Cuba]. / Diseño e instauración de un banco nacional de sueros para la vigilancia de enfermedades exóticas de los animales en la República de Cuba.

A serum bank for the surveillance of exotic diseases was designed in accordance with the provisions of the Information and Epizootiological Surveillance System in the Republic of Cuba. Sera were collected from imported animals, from sentinel animals used for monitoring target areas at biological risk and from animals located in high animal-density areas. Methodologies were developed for the selection and characterisation of target areas at biological risk and sentinel animal points, the collection and storage of serum samples and the management of the national animal serum bank. After developing the methodologies, the serum bank was established throughout Cuba. The national animal serum bank operates using a quality management system based on the recommendations of the World Organisation for Animal Health and the International Organization for Standardization.

Hematologic, cytochemical, ultrastructural, and molecular findings of Hepatozoon-infected flat-headed cats (Prionailurus planiceps).

BACKGROUND: The flat-headed cat (Prionailurus planiceps) is a small wild cat of Southeast Asia and is considered extremely endangered. Little is known about the hematologic values, blood cell morphology, or hemoparasites of this species in relation to other Felidae. OBJECTIVES: The objective of this study was to report basic hematologic values and describe the light microscopic, cytochemical, and ultrastructural characteristics of blood cells in 2 wild-caught flat-headed cats. In addition, molecular analysis was done of a Hepatozoon organism found in the neutrophils of both cats. METHODS: Blood samples were collected into EDTA from the cephalic vein. A CBC, manual differential count, manual reticulocyte count, cytochemical stains (Sudan black B [SBB], alpha-naphthyl acetate esterase [ANAE], and beta-glucuronidase), and scanning and transmission electron microscopy were done using standard methods. RESULTS: HCT was slightly lower and reticulocyte counts and red cell distribution width were higher than the expected values for other species of cats. Hepatozoon organisms were found in the cytoplasm of neutrophils in both cats, but the number of infected neutrophils was very low (1%-2%). Neutrophils stained strongly positive for SBB, but were negative for ANAE and beta-glucuronidase. Hepatozoon-infected neutrophils were negative for SBB, but focally positive for ANAE and beta-glucuronidase. By transmission electron microscopy, gamonts of Hepatozoon sp were observed in neutrophils, and rarely free in plasma. Infected neutrophils had fewer specific granules and more mitochondria compared with noninfected neutrophils. PCR products of partial 18S rRNA revealed that the isolate of Hepatozoon in the flat-headed cats was closely related to that of the frog Hepatozoon sp. CONCLUSIONS: These results add to our understanding of hematologic values and blood cell morphology in Hepatozoon-infected flat-headed cats as well as the molecular analysis of the Hepatozoon organism, and may be useful for the health management and evaluation of hemoparasitic disease in this species.

Sample Collection.

Results may be changed with suboptimal sample collection and transport, which then result in incorrect diagnoses. Quality management of samples must start in the patient, extend through sampling itself, include appropriate short transport, and then be correctly accessioned at the referral laboratory or in-house station to ensure accurate diagnosis of disease. A quality assurance plan at the laboratory chosen is mandatory under American Society for Veterinary Clinical Pathology guidelines.

Investigation of multiple mortality events in eastern box turtles (Terrapene carolina carolina).

Wildlife mortality investigations are important for conservation, food safety, and public health; but they are infrequently reported for cryptic chelonian species. Eastern box turtles (Terrapene carolina carolina) are declining due to anthropogenic factors and disease, and while mortality investigations have been reported for captive and translocated individuals, few descriptions exist for free-living populations. We report the results of four natural mortality event investigations conducted during routine health surveillance of three Illinois box turtle populations in 2011, 2013, 2014, and 2015. In April 2011, over 50 box turtles were found dead and a polymicrobial necrotizing bacterial infection was diagnosed in five survivors using histopathology and aerobic/anaerobic culture. This represents the first reported occurrence of necrotizing bacterial infection in box turtles. In August 2013, paired histopathology and qPCR ranavirus detection in nine turtles was significantly associated with occupation of moist microhabitats, identification of oral plaques and nasal discharge on physical exam, and increases in the heterophil count and heterophil to lymphocyte ratio (p < 0.05). In July 2014 and 2015, ranavirus outbreaks reoccurred within a 0.2km radius of highly-disturbed habitat containing ephemeral ponds used by amphibians for breeding. qPCR ranavirus detection in five individuals each year was significantly associated with use of moist microhabitats (p < 0.05). Detection of single and co-pathogens (Terrapene herpesvirus 1, adenovirus, and Mycoplasma sp.) was common before, during, and after mortality events, but improved sample size would be necessary to determine the impacts of these pathogens on the occurrence and outcome of mortality events. This study provides novel information about the causes and predictors of natural box turtle mortality events. Continued investigation of health, disease, and death in free-living box turtles will improve baseline knowledge of morbidity and mortality, identify threats to survival, and promote the formation of effective conservation strategies.

Acute Phase Protein Levels as An Auxiliary Tool in Diagnosing Viral Diseases in Ruminants-A Review.

We examined acute phase protein (APP) concentrations in viral infections of dairy ruminants and assessed the potential role of characteristic patterns of APP changes in auxiliary diagnosing viral diseases. All viruses reviewed are common causes of farm animal diseases. APPs are among the first agents of immunity, and their concentrations could be diagnostically relevant. In the most common ruminant viral diseases, elevated serum amyloid A (SAA) and haptoglobin (Hp) levels in blood serum have been observed. However, since these proteins are the main APPs in many viral infections, it is impossible to use their levels for diagnosing particular infections. Decreased Cp and albumin expression could help differentiate the bluetongue virus infection from other diseases. Lastly, analysis of SAA levels in blood serum and milk could be helpful in diagnosing small ruminant lentivirus infection. While promising, APP levels can only be considered as an auxiliary tool in diagnosing viral diseases in ruminants.

Clinical, virological and epidemiological characterization of an outbreak of Testudinid Herpesvirus 3 in a chelonian captive breeding facility: Lessons learned and first evidence of TeHV3 vertical transmission.

Testudinid herpesviruses (TeHVs) have a worldwide distribution among tortoises. However, information such as risk factors promoting the occurrence or the recrudescence of the associated disease and the mid-term sequelae of an outbreak comprising the extent and dynamic of the viral shedding have been only minimally investigated. Critical management information is also lacking or anecdotal. Furthermore, major aspects of the viral pathogenesis including the likelihood of vertical transmission of the virus are virtually unknown. The present study describes the occurrence and the management of an outbreak of Testudinid herpesvirus genotype 3 (TeHV3) in a large, private collection of chelonians. Clinical, pathological, molecular and serological characterization of the outbreak were carried out. Seventy-five percent of the infected tortoises died. Complementation of molecular and serological testing was a critical point for successful management implementations. A case-control study was performed to analyze possible risk factors associated with the infection. Furthermore, a subgroup of six asymptomatic infected tortoises was monitored for two consecutive seasons after the outbreak: all the infected tortoises were determined to be intermittent shedders, except one, which was a persistent shedder. Post-hibernation was associated with the highest number of shedders. Finally, evidence of the most likely vertical transmission of the virus was obtained for the first time.

Sero-epidemiology and hemato-biochemical study of bovine leptospirosis in flood affected zone of Pakistan.

The bovine leptospirosis is an economically important zoonotic disease of flood affected areas worldwide, but scarce information is available about its epidemiology in Pakistan. This is a first study on sero-epidemiology of bovine leptospirosis in Pakistan. The objectives of this study were to investigate the sero-prevalence and associated risk factors of bovine leptospirosis in flood affected zone of Punjab, Pakistan. A total of 385 serum samples were randomly collected from four tehsils of district Muzaffargarh, Pakistan. The serum samples were subjected to indirect ELISA for the detection of anti-leptospira antibodies. The overall sero-prevalence of leptospirosis was 30.39%. The prevalence was significantly higher (p<0.001) in cattle (56.25%) than buffaloes (4.66%). The key risk factors identified based on multivariable logistic regression were; confined system of rearing, flooded area, and lesser to graduate level of education as risk factors for leptospirosis. The values of hematological parameters varied significantly (p<0.05) for Hb, TEC and PCV while there was no significant (p>0.05) difference among TLC values among sero-positive and sero-negative animals. The serum biochemical profile revealed significant differences (p<0.05) in values of ALT, AST and creatinine among the sero-positive and negative animals. Hence, it can be concluded that leptospirosis is an emerging and neglected disease in flood affected zone of Pakistan, and the disease needs to be explored comprehensively in other parts of the country to sort out solid strategies for its control and eradication.

Tuberculosis outbreak in a dromedary racing herd and rapid serological detection of infected camels.

A recent outbreak of tuberculosis (TB) in a dromedary racing herd of 58 animals involved 3 infected animals. Disease was confirmed at necropsy by finding gross lesions from which Mycobacterium bovis (antelope type) was isolated. Sera collected from the camels in this herd were used to evaluate two new serological methods, Multiantigen Print Immunoassay (MAPIA) and rapid test (RT) developed using the lateral-flow technology, in comparison with the intradermal tuberculin tests. Antibodies were found in all three infected dromedaries by both RT and MAPIA, but not in the remaining 55 animals in the herd. With the limited number of animals tested in this study, the serological assays showed the potential for convenient, rapid, and accurate diagnosis of TB in live camels.

Serum concentration of copper, zinc, iron, and cobalt and the copper/zinc ratio in horses with equine herpesvirus-1.

The serum concentrations of copper, zinc, iron, and cobalt and copper/zinc ratio were investigated in horses infected with equine herpesvirus-1 (EHV-1). Nine horses were naturally infected with the virus and nine healthy horses served as controls. The concentrations of copper, zinc, iron, and cobalt were determined spectrophotometrically in the blood serum of all horses. The results were (expressed in micrograms per deciliters) copper 2.80 +/- 0.34 vs 1.12 +/- 0.44, zinc 3.05 +/- 0.18 vs 0.83 +/- 0.06, iron 2.76 +/- 0.17 vs 3.71 +/- 0.69, cobalt 0.19 +/- 0.37 vs 0.22 +/- 0.45, and copper/zinc ratio 0.72 +/- 0.38 vs 1.41 +/- 0.36 for control vs infected group, respectively. In conclusion, copper and zinc concentrations of the infected group were lower than the control group (p < 0.001), whereas iron concentration and the copper/zinc ratio of the infected group were higher than the control group (p < 0.05 and p < 0.001). The cobalt concentration was not found to be statistically different between two groups. It might be emphasized that copper/zinc ratio was significantly affected by the EHV-1 infection, so it could be taken into consideration during the course of infection.

Seroprevalence of leptospirosis in Korean municipal zoo animals.

From 2002 to 2005, we collected 118 serum samples from 34 species belonging to 13 families of zoo animals in Korea and determined the prevalence of antibodies for 18 serovars of Leptospira spp. using the microscopic agglutination test (MAT). Twenty-nine (25%) of the serum samples tested were positive for one or more of the serovars. There were no significant differences in relation to genders: 23% and 26% of positives occurring in male and female animals, respectively (P>0.05). However, the seroprevalence for the Leptospira spp. was significantly higher (P<0.05) in herbivores (45%) than in either carnivores (17%) or omnivores (17%). Among the 5 serovars detected in this study, the most common was sejroe (n=27; 87% of all positive reactions). All positive reactions showed low titers (< or = 1:200) and the positives were most frequently detected in 1:25 (58%) and 1:50 (23%) serum dilutions. The highest antibody titer (1:200) was observed for the serovars sejroe (n=1) and bratislava (n=1). We conclude that the exposure of zoo animals to Leptospira spp. is relatively common in Korea and produces low MAT titers, with sejroe being the most commonly encountered serovar.

Some morphological, biochemical and immunological blood parameters in European bison with or without lesions in the digestive, respiratory, urinary and reproductive tracts.

Ninety-seven bison (39 males and 58 females) were culled out during three successive winter seasons. No infection with BHV-1 and BHV-5 or other viruses was detected by PCR and SN methods. Advanced pathological lesions observed in males exhibiting symptoms of posthitis/balanoposthitis are probably due to secondary infection by microorganisms present in the prepuce (Corynebacterium spp., Bacillus spp., Fusobacterium necrophorum, Staphylococcus spp. (coagulase negative) and Staphylococcus hyicus). Histopathological examinations of sections from prepuce and lips of pudendum showed the presence of Onchocercinae nematode (Onchocerca sp.). A comparison of biochemical and morphological blood parameters in the bison with or without lesions in the digestive (presence of parasites), respiratory, urinary and reproductive tracts revealed that the most variable parameter during reproductive organ disorder is the leukocyte count, while disorders in the urinary and digestive tracts coincided with the age of animals. In cases of digestive disorder most changes were observed in the number of neutrophils and concentration of urea. The number of red blood cells, monocytes, the level of alkaline phosphatase (AP) and hematocrit negatively correlated with age. Negative correlation was also observed between the number of neutrophils, level of creatinine and lymphocyte proliferation index. However, positive correlation was seen between erythrocyte sedimentation rate (Spearman's correlation coefficient), level of alanine aminotransferase (ALT) and protein, and age of the bison. Positive correlation was also observed between the lymphocyte count and lymphocyte proliferation index. The attempt to determine the reference values for blood biochemical and morphological parameters showed similarities to those of cattle.

Seroprevalence of Q fever among human and animal in Iran; A systematic review and meta-analysis.

BACKGROUND: Q fever is a main zoonotic disease around the world. The aim of this meta-analysis was to estimate the overall seroprevalence of Coxiella burnetii among human and animal population in Iran. METHODS: Major national and international databases were searched from 2005 up to August 2016. We extracted the prevalence of Q fever antibodies (IgG) as the main primary outcome. We reported the prevalence of the seropositivity as point and 95% confidence intervals. RESULTS: The overall seroprevalence of IgG phase I and II antibodies of Q fever in human was 19.80% (95% CI: 16.35-23.25%) and 32.86% (95% CI: 23.80-41.92%), respectively. The herd and individual prevalence of C. burnetii antibody in goat were 93.42% (95% CI: 80.23-100.00) and 31.97% (95% CI: 20.96-42.98%), respectively. The herd and individual prevalence of Q fever antibody in sheep's were 96.07% (95% CI: 89.11-100.00%) and 24.66% (95% CI: 19.81-29.51%), respectively. The herd and individual prevalence of C. burnetii antibody in cattle were 41.37% (95% CI: 17.88-64.86%) and 13.30% (95% CI: 2.98-23.62%), respectively. Individual seropositivity of Q fever in camel and dog were 28.26% (95% CI: 21.47-35.05) and 0.55% (0.03-2.68), respectively. CONCLUSION: Seroprevalence of Q fever among human and domestic animals is considerable. Preventative planning and control of C. burnetii infections in Iran is necessary. Active surveillance and further research studies are recommended, to more clearly define the epidemiology and importance of C. burnetii infections in animals and people in Iran.

Clinical and Pathologic Features of a Suspected Selenium Deficiency in Captive Plains Zebras.

Previous studies have shown that selenium (Se) deficiency is associated with nutritional myopathy, known as white muscle disease (WMD), in horses. However, correlations between Se deficiency and clinical findings, such as hematologic biochemical values and pathological features, have not been evaluated in captive plains zebras. The purpose of the present study was to investigate the clinical and pathologic features that may be caused by a Se deficiency in the captive plains zebra. Clinical findings, feed analyses, hematologic biochemical analyses, response to treatment, and pathologic examination were assessed in six affected plains zebras. The dietary concentration of Se in feed was also tested. Sudden death occurred in two cases during the first day of the onset of symptoms. Two zebras died at 4 days and two zebras survived after treatment. The clinical signs in affected animals were characterized by general weakness, astasia, and abnormal postural positions. The Se concentration in hay from the breeding stable was low, based on the reference value. Glutathione peroxidase (GSH-Px) activity was lower compared with the equine reference value. Multiple areas of subcutaneous steatitis and pale skeletal muscle and myocardium were revealed at gross necropsy. Degeneration and necrosis of myocardial and skeletal muscles, as well as congestion of the liver, lung, and kidney were found via histopathological examination. No suspected bacterial infections were found. Feed analyses, response to treatment, serum GSH-Px activity, and pathological features suggest that Se deficiency may have caused the disease in the six affected captive plains zebra.