Transcriptome-level assessment of the impact of deformed wing virus on honey bee larvae.

Deformed wing virus (DWV) prevalence is high in honey bee (Apis mellifera) populations. The virus infects honey bees through vertical and horizontal transmission, leading to behavioural changes, wing deformity, and early mortality. To better understand the impacts of viral infection in the larval stage of honey bees, artificially reared honey bee larvae were infected with DWV (1.55 × 1010 copies/per larva). No significant mortality occurred in infected honey bee larvae, while the survival rates decreased significantly at the pupal stage. Examination of DWV replication revealed that viral replication began at 2 days post inoculation (d.p.i.), increased dramatically to 4 d.p.i., and then continuously increased in the pupal stage. To better understand the impact of DWV on the larval stage, DWV-infected and control groups were subjected to transcriptomic analysis at 4 d.p.i. Two hundred fifty-five differentially expressed genes (DEGs) (fold change ≥ 2 or ≤ -2) were identified. Of these DEGs, 168 genes were downregulated, and 87 genes were upregulated. Gene Ontology (GO) analysis showed that 141 DEGs (55.3%) were categorized into molecular functions, cellular components and biological processes. One hundred eleven genes (38 upregulated and 73 downregulated) were annotated by KO (KEGG Orthology) pathway mapping and involved metabolic pathways, biosynthesis of secondary metabolites and glycine, serine and threonine metabolism pathways. Validation of DEGs was performed, and the related gene expression levels showed a similar tendency to the DEG predictions at 4 d.p.i.; cell wall integrity and stress response component 1 (wsc1), cuticular protein and myo-inositol 2-dehydrogenase (iolG) were significantly upregulated, and small conductance calcium-activated potassium channel protein (SK) was significantly downregulated at 4 d.p.i. Related gene expression levels at different d.p.i. revealed that these DEGs were significantly regulated from the larval stage to the pupal stage, indicating the potential impacts of gene expression levels from the larval to the pupal stages. Taken together, DWV infection in the honey bee larval stage potentially influences the gene expression levels from larvae to pupae and reduces the survival rate of the pupal stage. This information emphasizes the consequences of DWV prevalence in honey bee larvae for apiculture.

MORBIDITY AND MORTALITY OF TAKIN (BUDORCAS TAXICOLOR) IN NORTH AMERICAN ZOOLOGICAL INSTITUTIONS FROM 1997 TO 2017.

Takin (Budorcus taxicolor) are classified as "Vulnerable" on the International Union for the Conservation of Nature Red List. Thus, ex situ conservation efforts provide assurance populations for future survival of this species. The objective of this study was to identify common causes of morbidity and mortality in takin populations in human care. Twenty North American institutions that housed takin from 1997 to 2017 completed a survey requesting medical and husbandry data. Data were examined broadly, by sex and age groups. There were 206 morbidity events (male = 133; female = 73) submitted across 102 takin (male = 62; female = 40). The most common causes of morbidity were infectious or inflammatory diseases (50%; 104/206), degenerative diseases (22%; 46/206), and traumatic events (17%; 34/206). Necropsy reports were provided for 42 takin that died during the study period. The most common causes of mortality were infectious or inflammatory diseases (26%; 11/42), traumatic events (24%; 10/42), and degenerative disease (12%; 5/42). Sixty-two percent of infectious or inflammatory diseases causing morbidity were associated with endoparasites (64/104). Degenerative joint diseases more commonly affected males (78%; 36/46) as well as forelimbs (48%; 22/46) when compared to hindlimbs (30%; 14/46) and unspecified limbs (22%; 10/46). The prevalence of trauma as a cause of morbidity and mortality was higher in neonate and juvenile takin groups combined (morbidity = 19%; mortality = 50%) as compared to adult and senior takin groups combined (morbidity = 15%; mortality = 11%). Older takin were euthanatized more often (57%; 16/28) than younger takin (29%; 4/14). Correlations between husbandry and health were difficult as a result of the inherent limitations of the survey. These data will inform takin-holding zoologic institutions and contribute to the successful management of takin in human care.

Continued preference for suboptimal habitat reduces bat survival with white-nose syndrome.

Habitat alteration can influence suitability, creating ecological traps where habitat preference and fitness are mismatched. Despite their importance, ecological traps are notoriously difficult to identify and their impact on host-pathogen dynamics remains largely unexplored. Here we assess individual bat survival and habitat preferences in the midwestern United States before, during, and after the invasion of the fungal pathogen that causes white-nose syndrome. Despite strong selection pressures, most hosts continued to select habitats where disease severity was highest and survival was lowest, causing continued population declines. However, some individuals used refugia where survival was higher. Over time, a higher proportion of the total population used refugia than before pathogen arrival. Our results demonstrate that host preferences for habitats with high disease-induced mortality can create ecological traps that threaten populations, even in the presence of accessible refugia.

Laboratory Infection of Novel Akhmeta Virus in CAST/EiJ Mice.

Akhmeta virus is a zoonotic Orthopoxvirus first identified in 2013 in the country of Georgia. Subsequent ecological investigations in Georgia have found evidence that this virus is widespread in its geographic distribution within the country and in its host-range, with rodents likely involved in its circulation in the wild. Yet, little is known about the pathogenicity of this virus in rodents. We conducted the first laboratory infection of Akhmeta virus in CAST/EiJ Mus musculus to further characterize this novel virus. We found a dose-dependent effect on mortality and weight loss (p < 0.05). Anti-orthopoxvirus antibodies were detected in the second- and third-highest dose groups (5 × 104 pfu and 3 × 102 pfu) at euthanasia by day 10, and day 14 post-infection, respectively. Anti-orthopoxvirus antibodies were not detected in the highest dose group (3 × 106 pfu), which were euthanized at day 7 post-infection and had high viral load in tissues, suggesting they succumbed to disease prior to mounting an effective immune response. In order of highest burden, viable virus was detected in the nostril, lung, tail, liver and spleen. All individuals tested in the highest dose groups were DNAemic. Akhmeta virus was highly pathogenic in CAST/EiJ Mus musculus, causing 100% mortality when ≥3 × 102 pfu was administered.

An integrated omics approach to investigate summer mortality of New Zealand Greenshell™ mussels.

BACKGROUND: Green-lipped mussels, commercially known as Greenshell™ mussels (Perna canaliculus Gmelin 1791), contribute > $300 million to New Zealand's aquaculture exports. However, mortalities during summer months and potential pathogenic outbreaks threaten the industry. Thermal stress mechanisms and immunological responses to pathogen infections need to be understood to develop health assessment strategies and early warning systems. METHODS: P. canaliculus were collected during a mortality event at a commercial aquaculture farm in Firth of Thames, New Zealand. Gill tissues from six healthy and six unhealthy mussels were excised and processed for metabolomic (GC-MS) and label-free proteomic (LC-MS) profiling. Univariate analyses were conducted separately on each data layer, with data being integrated via sparse multiple discriminative canonical correlation analysis. Pathway enrichment analysis was used to probe coordinated changes in functionally related metabolite sets. RESULTS: Findings revealed disruptions of the tricarboxylic acid (TCA) cycle and fatty acid metabolism in unhealthy mussels. Metabolomics analyses also indicated oxidative stress in unhealthy mussels. Proteomics analyses identified under-expression of proteins associated with cytoskeleton structure and regulation of cilia/flagellum in gill tissues of unhealthy mussels. Integrated omics revealed a positive correlation between Annexin A4 and CCDC 150 and saturated fatty acids, as well as a negative correlation between 2-aminoadipic acid and multiple cytoskeletal proteins. CONCLUSIONS: Our study demonstrates the ability of using integrative omics to reveal metabolic perturbations and protein structural changes in the gill tissues of stressed P. canaliculus and provides new insight into metabolite and protein interactions associated with incidences of summer mortality in this species.

A unifying framework for the transient parasite dynamics of migratory hosts.

Migrations allow animals to track seasonal changes in resources, find mates, and avoid harsh climates, but these regular, long-distance movements also have implications for parasite dynamics and animal health. Migratory animals have been dubbed "superspreaders" of infection, but migration can also reduce parasite burdens within host populations via migratory escape from contaminated habitats and transmission hotspots, migratory recovery due to parasite mortality, and migratory culling of infected individuals. Here, we show that a single migratory host-macroparasite model can give rise to these different phenomena under different parametrizations, providing a unifying framework for a mechanistic understanding of the parasite dynamics of migratory animals. Importantly, our model includes the impact of parasite burden on host movement capability during migration, which can lead to "parasite-induced migratory stalling" due to a positive feedback between increasing parasite burdens and reduced movement. Our results provide general insight into the conditions leading to different health outcomes in migratory wildlife. Our approach lays the foundation for tactical models that can help understand, predict, and mitigate future changes of disease risk in migratory wildlife that may arise from shifting migratory patterns, loss of migratory behavior, or climate effects on parasite development, mortality, and transmission.

A RETROSPECTIVE SUMMARY OF CERVID MORBIDITY AND MORTALITY IN ONTARIO AND NUNAVUT REGIONS OF CANADA (1991-2017).

Free-ranging cervids in Canada face diverse threats such as climate change, human population expansion, and the northward spread of vector-borne pathogens. However, we currently have a limited understanding of the impacts of these health challenges in Ontario cervids. Our objective was to identify and characterize causes of morbidity and mortality in free-ranging cervids submitted to the Ontario and Nunavut node of the Canadian Wildlife Health Cooperative (CWHC) over a 27-yr period (1991 to 2017). Submissions included carcasses submitted for full postmortem examination (gross and histopathology; n=196) and field-collected tissues (n=384). Ancillary tests were performed on a case-by-case basis. Univariable logistic regression was used to test for associations between select causes of morbidity and mortality, and factors such as sex, age, and season. Four cervid species were examined: white-tailed deer (Odocoileus virginianus; n=211), moose (Alces alces; n=140), elk (Cervus canadensis; n=136), and caribou (Rangifer tarandus caribou; n=93). Noninfectious disease was the most common general cause of morbidity and mortality (38.1%; 221/580) and was most commonly attributed to trauma (49.7%; 110/221). Deaths attributed to infectious diseases (34.3%; 199/580) were most often bacterial in etiology (45.7%; 91/199). The most common primary infectious disease diagnosed in caribou was digital limb infection, and moose were most commonly diagnosed with parasitic causes, including meningeal worm (Parelaphostrongylus tenuis) and winter tick (Dermacentor albipictus). Chronic wasting disease was not diagnosed among cervids tested during the study period, consistent with the previous lack of detection of the disease in Ontario. These results reveal that anthropogenic, infectious, and environmentally associated causes of morbidity and mortality are commonly diagnosed in cervids submitted to the CWHC Ontario and Nunavut regions, and represent potential population threats that should continue to be monitored.

CLOUDED LEOPARD (NEOFELIS NEBULOSA) MORBIDITY AND MORTALITY IN CAPTIVE-BRED POPULATIONS: A COMPREHENSIVE RETROSPECTIVE STUDY OF MEDICAL DATA FROM 271 INDIVIDUALS IN EUROPEAN, ASIAN, AND AUSTRALIAN ZOOS.

The clouded leopard (Neofelis nebulosa) is classified as vulnerable on the International Union for the Conservation of Nature Red List of Threatened Species. However, diseases affecting this species across zoo populations are not well documented. The primary objective of this retrospective study was to identify common and significant causes of morbidity and mortality in captive-bred clouded leopards from European, Asian, and Australian institutions. Medical records from 44 zoological parks that held 271 clouded leopards from 1934 to 2017 were reviewed. Major causes of mortality in the dead leopards (n = 141) were respiratory disease (17%), maternal neglect and starvation (12%), generalized infectious disease (10%), digestive disease (10%), and trauma (10%). Six animals lived more than 20 yr and two were older than 22 yr. Diseases were recorded 344 times (average of two per leopard) in 166 living leopards. The body systems most frequently affected by disease in these 166 individuals were, in order of frequency, integumentary (prevalence = 21%), digestive (21%), respiratory (16%), musculoskeletal (12%), and urinary (10%) systems. Neoplasia (7%) was less frequent, followed by cardiovascular (5%), genital (3%), and viral (3%) disorders. Extensive, self-induced alopecia on the tail and dorsum was the most frequently reported dermatological disease, which is proposed to be called the "clouded leopard alopecia syndrome." The most common neoplasm was pheochromocytoma (1%), followed by squamous cell carcinoma of the paw pads, pleural mesothelioma and multicentric lymphomas (<1% each). Dilated cardiomyopathy (2%) was the most common cardiovascular disease. Bronchopneumonia (7%), enteritis (4%), and nephritis (4%) were the most frequently reported respiratory, digestive, and renal diseases, respectively. Diagnosed disease incidence was significantly higher in Europe. This paper reports the results of a comprehensive study of the causes of morbidity and mortality in European, Asian, and Australian clouded leopard zoo populations.

RETROSPECTIVE EVALUATION OF THE USE OF PARENTERAL NUTRITION IN HOSPITALIZED NONDOMESTIC RUMINANTS.

Parenteral nutrition (PN) is one method of providing nutrient support to hospitalized, nondomestic ruminants that have a decreased appetite in hospital or have high metabolic demands caused by illness. There are a limited number of published reports of the use of PN in nondomestic ruminants. A retrospective evaluation of PN use in adult (>6 mo of age) hospitalized ruminants at the San Diego Zoo Safari Park was conducted from 2014 to 2018 (n = 24). Discharge rate for animals that received PN was 34%. Poor survival was likely caused by case selection of animals that had severe disease or malnutrition necessitating the need for PN. Common metabolic changes among the study animals included the following: hypokalemia, hyperglycemia, and hyperphosphatemia or hypophosphatemia. Bivariable analysis revealed no clinically significant factors that influenced odds of survival. There was little evidence of adverse effects with the administration of PN during the study period. Parenteral nutrition requires specialized equipment and technical skills, but is a viable means of nutrient support for hospitalized nondomestic ruminants.

PROGNOSTIC INDICATORS FOR SURVIVAL OF ORPHANED NEONATAL AND JUVENILE EASTERN COTTONTAIL RABBITS (SYLVILAGUS FLORIDANUS): 1,256 CASES (2012-17).

Orphaned eastern cottontail rabbits (ECRs; Sylvilagus floridanus) often present to wildlife clinics within their geographic range and require considerable dedication of time and resources. The objective of this analytical cross-sectional study was to assess initial examination findings to be used as prognostic indicators for orphaned neonatal and juvenile ECRs. The medical records of the University of Illinois Wildlife Medical Clinic were searched for ECRs presenting between 2012 and 2018. This criterion identified 1,256 ECRs that were then classified as survivors (survived and released) or as nonsurvivors (euthanized or natural death) within 72 h of admission. Presenting weight, body system abnormalities, hydration status, intervention prior to presentation, and singleton versus group presentation were categorically recorded for each individual ECR. The data were modeled using a series of logistic regression models fitted using the general linear model. Individuals were significantly more likely to be nonsurvivors if they presented as singletons (P<0.0001), presented with moderate/severe (P<0.001) or mild integumentary signs (P=0.0261), presented with multi-organ disease (P<0.001), presented with neurologic signs (P<0.0003), or had treatment provided prior to presentation (P=0.031). Factors that did not predict survival status in ECRs included body weight (P=0.210), presence of respiratory signs (P=0.674), and presence of dehydration (P=0.356). These findings may be used at wildlife medical clinics to make triage criteria for euthanasia as well as dedicate limited funds and labor to cases with the best prognosis for survival.

RETROSPECTIVE STUDY OF CAUSES OF ANIMAL MORTALITY IN LJUBLJANA ZOO 2005-2015.

Zoos harbor large collections of diverse species, aiding in both conservation and education, as well as research in multiple scientific fields. However, the most common causes of death in zoo animals around the world remain unclear because few extensive reports or reviews are published on this topic. This information could greatly improve preventive veterinary medicine in zoologic gardens. This study provides a retrospective overview of the causes of death of animals from the Ljubljana Zoo in the years 2005-2015. During this period, a total of 353 animals were submitted for necropsy, of which 244 were mammals, 85 were birds, and 25 were reptiles. The causes of deaths were divided into infectious diseases (38%), dysfunctions of individual organs (20%), traumas (13%), parasitosis (7%), reproductive disorders (6%), metabolic disorders (3%), neoplastic disease (4%), and intoxications (4%). In some cases, the cause of death was unable to be determined (7%), most commonly because of autolysis of the body. The results of this retrospective study bring a general overview of the epizootiologic situation in the Ljubljana Zoo over an 11-yr period and valuable information to other zoos to optimize preventative plans and diagnostics.

Morbidity and mortality of domestic rabbits (Oryctolagus cuniculus) under primary veterinary care in England.

BACKGROUND: The domestic rabbit is a common pet species, but limited research exists on the health of pet rabbits. This study aimed to characterise common disorders of pet rabbits and reasons for mortality as recorded by veterinary practices in England. METHODS: This cross-sectional study covered anonymised clinical records of 6349 rabbits attending 107 primary veterinary care clinics. RESULTS: The median age was 3.2 years (interquartile range (IQR) 1.6-5.1), and the median adult bodyweight was 2.1 kg (IQR 1.7-2.6). The most common breed types were domestic (n=2022, 31.9 per cent), lop (1675, 26.4 per cent) and Netherland dwarf (672, 10.6 per cent). For those rabbits that died during the study period, the median age at death was 4.3 years (IQR 2.1-7.0). The most common causes of death were recorded as myiasis (prevalence 10.9 per cent, 95 per cent confidence interval (CI): 7.4 to 15.2), anorexia (4.9 per cent, 95 per cent CI: 4.0 to 10.4), recumbency/collapse (4.9 per cent, 95 per cent CI: 4.0 to 10.4) and ileus (4.3 per cent, 95 per cent CI: 3.5 to 9.5). The most prevalent specific disorders recorded were overgrown claw/nails (16.0 per cent, 95 per cent CI: 14.5 to 17.5), overgrown molar(s) (7.6 per cent, 95 per cent CI: 6.6 to 8.7), perineal soiling (4.5 per cent, 95 per cent CI: 3.7 to 5.4), overgrown incisor(s) (4.3 per cent, 95 per cent CI: 3.5 to 5.2) and ileus (4.2 per cent, 95 per cent CI: 3.4 to 5.0). CONCLUSIONS: This study augments the limited evidence base on rabbit health and can assist veterinarians to better advise owners on optimal animal husbandry priorities.

MORTALITY INVESTIGATION OF MONITORED EASTERN BOX TURTLES (TERRAPENE CAROLINA CAROLINA) IN CENTRAL ILLINOIS, USA, FROM 2016-18.

Mortality events in eastern box turtles (Terrapene carolina carolina) threaten conservation efforts across the species range. These events are often under-diagnosed and, when observed, predictive health factors are unavailable prior to death. At Kickapoo State Park in central Illinois, USA, ranaviruses caused observed mortality events in amphibians and chelonians in 2014 and 2015. Following these outbreaks, eastern box turtles (n=36) were affixed with radio transmitters and temperature data loggers to obtain repeated location and temperature data from spring 2016-spring 2018. Bimonthly, samples of blood and oral and cloacal swabs were collected to investigate health parameters (hematology and cytokine transcription) and presence of multiple pathogens. Deaths of instrumented turtles occurred in 2016 (n=5), 2017 (n=15), and 2018 (n=2). The largest single die-off occurred in February 2017 (n=7). Seventeen turtles were necropsied and multiple pathologic processes were identified, most frequently decreased adipose stores (n=6). Two turtles had pathologic findings consistent with multisystemic inflammation. In addition, infectious pathogens were identified in turtles prior to death, but no single agent was associated with each mortality event. Ranavirus was not detected in any turtle. Hot spot analysis revealed spatial clustering at the center and edges of the study area for body temperature as well as for relative cytokine transcription of interleukin-1 beta, tumor necrosis factor alpha, and interleukin-10 associated with turtle death. Though no single causal factor could be identified, the information from this mortality event can direct future chelonian mortality investigations by providing baseline longitudinal data prior to death and in surviving turtles.

A molecular clone of Chronic Bee Paralysis Virus (CBPV) causes mortality in honey bee pupae (Apis mellifera).

Among the many diseases compromising the well-being of the honey bee (Apis mellifera) the chronic paralysis syndrome of adult honey bees is one of the best described. The causative agent, chronic bee paralysis virus (CBPV), is a positive sense, single-stranded RNA virus with a segmented genome. Segment 1 encodes three putative open reading frames (ORFs), including the RNA-dependent RNA polymerase and other non-structural protein coding regions. Segment 2 encodes four putative ORFs, which contain the genes of supposed structural proteins. In this study, we established a reverse genetic system for CBPV by molecular cloning of DNA copies of both genome segments. CBPV rescue was studied in imago and honey bee pupae infection models. Virus replication and progeny virus production was only initiated when capped RNAs of both genome segments were injected in honey bees. As injection of these clonal RNAs caused clinical symptoms similar to wild-type CBPV infection, we conclude that the novel molecular clone fulfilled Koch's postulates. Our virus clone will enable in-depth analysis of CBPV pathogenesis and help to increase knowledge about this important honey bee disease.

The Enhanced Immune Protection in Chinese Mitten Crab Eriocheir sinensis Against the Second Exposure to Bacteria Aeromonas hydrophila.

Accumulating evidences suggest that the enhanced immune responses and increased protection against bacteria-induced mortality can be initiated after the primary exposure to various microbial communities and their components in various organisms including commercially valuable crustaceans. In the present study, the survival rate and immune responses of Chinese mitten crab Eriocheir sinensis were determined after an immune priming (IP) with formalin-killed Aeromonas hydrophila and an immune challenge (ICH) with the same but live pathogen (Ah group). A group in which the animals received a salt injection prior to challenge was maintained as control (Ns group). In the present study, it was shown that an IP with killed A. hydrophila can significantly protect the crabs against the ICH with a lethal dose of the live pathogen. The increased survival was associated with elevated rate and duration of phagocytosis. The antibacterial activity of the serum was significantly increased in Ah group compared to that in Ns group. Significant changes of phenoloxidase (PO) activities were also found between Ah and Ns group but not in Ah group between IP and ICH. No significant changes of lysozyme were found in Ah and NS group during the whole experiment except 3 h after IP. In addition, the levels of transcripts and protein of Dscam were increased in hemocytes of the crabs from Ah group. All the results suggested that a primary immune priming with a particular killed pathogen could induce an enhanced immunity in crabs when they were encountered secondly with the same live pathogen. The evidences of elevated immune protections in crabs would contribute to better understand the mechanism of immune priming in invertebrates.

RETROSPECTIVE ANALYSIS OF MORTALITY IN CAPTIVE BONGO (TRAGELAPHUS EURYCERUS), 1995-2015.

A retrospective histologic study was performed for 96 deceased bongo (Tragelaphus eurycerus) submitted to Northwest ZooPath from 1995 to 2015. Histologic data were assessed for associations with sex, age, cause of death/euthanasia, and affected organ systems. Female bongo lived significantly longer than males. Males were more likely to die from infectious causes (41.2%), whereas most females died from chronic noninfectious conditions (54.4%) and trauma/stress (28.1%). Of those that died from infectious disease, the respiratory tract was the most commonly affected organ system. The most common infectious agents included acid-fast bacteria and fungi. Chronic conditions included amyloidosis (31.0%), inanition/emaciation (23.8%), and neoplasia (21.4%). Of the 31 animals that died with amyloidosis, 58% appeared to be clinically affected, and amyloidosis was likely an underlying cause of death in 42% of the animals. The most commonly affected organs included the liver, kidneys, adrenal glands, and gastrotintestinal tract. Also noteworthy was a high prevalence of adrenal gland hyperplasia and neoplasia, cystic thyroid glands, and aspiration pneumonia, which were not consistently associated with a prior anesthetic event or other obvious predisposing cause.

Increase in morbidity and mortality in a shipment of red-eared slider turtles (Trachemys scripta elegans).

A cohort of captive-bred red-eared slider turtles, Trachemys scripta, was received from a commercial vendor. Shortly after arrival, several turtles presented as lethargic with subjectively pale skin and multifocal areas of cotton-like tufts in the mouth area and distal extremities. The water was treated with a commercial anti-fungal and anti-bacterial preparation of Victoria Green B and acriflavine. Despite treatment, 10 turtles were euthanized and others demonstrated persistent clinical signs. A live turtle was submitted to a commercial diagnostic laboratory for microbiologic and histologic evaluation. Seven cultures were obtained from this turtle and numerous organisms grew from each culture, including Flavobacterium sp. Blood film analysis demonstrated intracytoplasmic gamonts of Haemogregarina sp. within erythrocytes. On necropsy, internal organs appeared to be slightly more adhered within the coelomic cavity than normal. The urinary bladder was markedly distended with turbid, dark yellow urine. Microscopic evaluation of the tissues revealed significant parasitism with Myxidium sp., Spirorchis sp. and Neopolystoma orbiculare. No fungal organisms were identified on histology or grown in culture. While there are scattered reports of these pathogens in freshwater turtles, none of the cases reported describe such extensive co-infections. It is likely that complicated infection and shipping stress exacerbated clinical signs typically seen with these organisms. Efforts to minimize stress and administration of prophylactic antiparasitic agents during the acclimation period may aid in reducing the consequences of internal parasitism in aquatic turtles.

Mass Die-Off of Saiga Antelopes, Kazakhstan, 2015.

In 2015, a mass die-off of ≈200,000 saiga antelopes in central Kazakhstan was caused by hemorrhagic septicemia attributable to the bacterium Pasteurella multocida serotype B. Previous analyses have indicated that environmental triggers associated with weather conditions, specifically air moisture and temperature in the region of the saiga antelope calving during the 10-day period running up to the event, were critical to the proliferation of latent bacteria and were comparable to conditions accompanying historically similar die-offs in the same areas. We investigated whether additional viral or bacterial pathogens could be detected in samples from affected animals using 3 different high-throughput sequencing approaches. We did not identify pathogens associated with commensal bacterial opportunisms in blood, kidney, or lung samples and thus concluded that P. multocida serotype B was the primary cause of the disease.