The golden jackal (Canis aureus): A new host for Echinococcus multilocularis and Trichinella britovi in Switzerland.

INTRODUCTION: The golden jackal (Canis aureus) is a wild canid new to Switzerland. It is an officially monitored species and all deceased individuals are submitted for post-mortem examination to collect baseline health data. This includes parasitological examinations, with an emphasis on zoonotic, reportable infections, such as those caused by Trichinella spp. or Echinococcus spp. From 2016 to 2021, five golden jackals originating from four Swiss cantons were submitted for full post-mortem examination. In one case only organ samples were available, and therefore parasitological examination was not possible. Parasite stages recovered during necropsy, as well as by routine coproscopical techniques, were morphologically identified. Taeniid eggs and adult tapeworms were processed for molecular species identification. Additionally, tongue and diaphragm were analysed for Trichinella spp. by the artificial digestion technique followed by multiplex-PCR in positive cases. Of the four jackals investigated for parasites, hookworm eggs were detected in one animal, both adult worms and eggs of Echinococcus multilocularis were present in another case, and one animal was free of parasites. Eggs of E. multilocularis as well as eggs of Toxocara canis and sporocysts of Sarcocystis sp. were detected in the intestinal content, and Trichinella britovi larvae were found in the muscle samples of the last case. The health monitoring programme in place for protected carnivores in Switzerland allowed us to add the golden jackal to the list of hosts for the endemic zoonotic parasites E. multilocularis and T. britovi in this country. Hunters, farmers, and other persons who could come in contact with golden jackals should be aware of the associated health risk and handle faeces and carcasses with caution.

INTRODUCTION: Le chacal doré (Canis aureus) est un canidé sauvage nouvellement présent en Suisse. Il s'agit d'une espèce officiellement surveillée et tous les individus morts sont soumis à un examen post-mortem afin de recueillir des données sanitaires de base. Cela inclut un examen parasitologique mettant l'accent sur les infections zoonotiques à déclaration obligatoire, telles que celles causées par Trichinella spp. ou Echinococcus spp. De 2016 à 2021, cinq chacals dorés originaires de quatre cantons suisses ont été soumis à un examen post-mortem complet. Dans un cas, seuls des échantillons d'organes ont été envoyés, l'examen parasitologique n'a pas été possible pour cet animal. Les stades parasitaires trouvés lors de l'examen pathologique et de la coprologie de routine ont été identifiés morphologiquement. Les espèces de ténias (œufs et stades adultes) ont été déterminées par des techniques de biologie moléculaire. En outre, la recherche de Trichinella spp. a été effectuée sur du tissu musculaire lingual et diaphragmatique par la technique de digestion artificielle suivie d'une PCR multiplex dans les cas positifs. Sur les quatre chacals ayant fait l'objet d'une recherche de parasites, des œufs d'ankylostomes ont été détectés chez un animal, des vers adultes et des œufs d'Echinococcus multilocularis étaient présents chez un autre animal, et aucun parasite n'a été trouvé dans un autre cas. Chez le dernier cas, des œufs d'E. multilocularis ainsi que des œufs de Toxocara canis et des sporocystes de Sarcocystis sp. ont été détectés dans le contenu intestinal, et des larves de Trichinella britovi ont été trouvées dans les échantillons de muscle. Le programme de surveillance sanitaire mis en place pour les carnivores protégés en Suisse a donc permis d'ajouter le chacal doré à la liste des hôtes des parasites zoonotiques endémiques E. multilocularis et T. britovi. Les chasseurs, agriculteurs et autres personnes susceptibles d'entrer en contact avec le chacal doré doivent être conscients du risque sanitaire associé et manipuler les fèces et les carcasses avec précaution.

Polycystic Kidney-like Disease in a Red-ear Slider Turtle (Trachemys scripta elegans).

A 14-year-old red-ear slider turtle (Trachemys scripta elegans) with no history of pre-existing clinical disease died and was referred for necropsy examination. Grossly, oesophageal prolapse, bilateral renal cysts and a paraduodenal cystic mass were detected. Tissues were processed routinely for histology and immunohistochemistry (IHC) with primary antibodies specific for Wilm's tumour suppressor gene-1 (WT-1), insulin, glucagon and pancytokeratins. Microscopically, renal medullary cysts and medullary atrophy resembled the changes associated with polycystic kidney disease (PKD). The cysts of the paraduodenal mass were lined by ciliated epithelial cells resembling embryonal cells and were intensely positive for glucagon and insulin by IHC. There was no cytokeratin expression in either lesion. WT-1 expression in the paraduodenal mass was cytoplasmic and appeared non-specific. Lesions were consistent with renal PKD-like disease and a pancreatic cystic hamartoma.

Causes of mortality and morbidity in free-ranging mustelids in Switzerland: necropsy data from over 50 years of general health surveillance.

BACKGROUND: Although mustelids occur worldwide and include a wide range of species, little is known about the diseases affecting them. Mustelids have regularly been submitted for post mortem investigation in the framework of the program for general wildlife health surveillance in Switzerland, which has been in place for nearly 60 years. We performed a retrospective analysis of the necropsy reports on mustelids submitted to the diagnostic service of the University of Bern. The aims of this study were to present an overview of the causes of mortality and morbidity observed in these carnivores, to assess differences among species, to assess changes in disease detection over the study period, and to describe the pathology of selected diseases. RESULTS: Five hundred and sixty-six reports from 1958 to 2015 were analyzed. Most animals were stone martens (Martes foina, 46%) and badgers (Meles meles, 44%); the remaining species were polecats (Mustela putorius, 4.7%), pine martens (Martes martes, 2%), stoats (Mustela erminea, 1.4%), weasels (Mustela nivalis, 0.8%) and otters (Lutra lutra, 0.3%). Infectious diseases (n = 262) were frequent and were mostly bacterial or viral; non-infectious conditions (n = 169) were less common and were mostly traumatic or due to metabolic disorders. The most frequent diagnoses included distemper (75% were badgers), amyloidosis (96% were martens), bacterial respiratory infections (all mustelids), biting lice (badgers only) and pulmonary and gastro-intestinal helminths (all species). Less frequent diseases included histoplasmosis (badgers only), aspergillosis, toxoplasmosis, hepatozoonosis, and sarcoptic mange. Lesions due to infection with distemper virus were primarily appreciated in the respiratory tract and central nervous system; they presented species-specific characteristics such as necrosis in the ependyma in badgers and absence of syncytia in stone martens. Amyloidosis in martens was multisystemic in most cases and included both AA and AL amyloidosis; the main macroscopic change was severe splenomegaly. CONCLUSION: Infectious diseases were the most frequent causes of morbidity and mortality of mustelids, with marked species-specific differences. Lung and skin were the most commonly affected organs. Contagious diseases such as canine distemper, sarcoptic mange and rabies in mustelids showed a similar temporal pattern as in red foxes (Vulpes vulpes), suggesting pathogen spillovers from foxes to mustelids.

Fatal leptospirosis in free-ranging Eurasian beavers (Castor fiber L.), Switzerland.

Leptospirosis was first diagnosed in free-ranging Eurasian beavers (Castor fiber L.) in Switzerland in 2010. Pathologic, serologic, molecular and epidemiologic analyses were carried out on 13 animals submitted for necropsy from 2010 through 2014. Typical lesions included alveolar haemorrhages in the lungs, tubular degeneration and interstitial nephritis in the kidneys. Microscopic agglutination test results were positive for serogroups Icterohaemorrhagiae, Australis, Autumnalis and Sejroe. Molecular analysis identified four distinct profiles belonging to serovar Icterohaemorrhagiae or Copenhageni. The severity and features of the lesions were consistent with a fatal disease associated with leptospires similarly to what has been reported in other animals and humans. The spatiotemporal occurrence of leptospirosis in beavers suggested an upstream spread of the bacteria and coincided with an increased incidence of leptospirosis in dogs and a case cluster in humans. However, an epidemiologic link among beaver cases and among species was not supported neither by the serologic nor molecular data.

Ranid Herpesvirus 3 and Proliferative Dermatitis in Free-Ranging Wild Common Frogs (Rana Temporaria).

Amphibian pathogens are of current interest as contributors to the global decline of amphibians. However, compared with chytrid fungi and ranaviruses, herpesviruses have received relatively little attention. Two ranid herpesviruses have been described: namely, Ranid herpesvirus 1 (RHV1) and Ranid herpesvirus 2 (RHV2). This article describes the discovery and partial characterization of a novel virus tentatively named Ranid herpesvirus 3 (RHV3), a candidate member of the genus Batrachovirus in the family Alloherpesviridae. RHV3 infection in wild common frogs (Rana temporaria) was associated with severe multifocal epidermal hyperplasia, dermal edema, a minor inflammatory response, and variable mucous gland degeneration. Intranuclear inclusions were numerous in the affected epidermis together with unique extracellular aggregates of herpesvirus-like particles. The RHV3-associated skin disease has features similar to those of a condition recognized in European frogs for the last 20 years and whose cause has remained elusive. The genome of RHV3 shares most of the features of the Alloherpesviruses. The characterization of this presumptive pathogen may be of value for amphibian conservation and for a better understanding of the biology of Alloherpesviruses.

Aeromonas salmonicida type III secretion system-effectors-mediated immune suppression in rainbow trout (Oncorhynchus mykiss).

Aeromonas salmonicida subsp. salmonicida, the etiologic agent of furunculosis, is a major pathogen in aquaculture. Together with other pathogens, it is characterized by the presence of a type 3 secretion system (T3SS). The T3SS is the main virulence mechanism of A. salmonicida. It is used by the bacterium to secrete and translocate several toxins and effector proteins into the host cell. Some of these factors have a detrimental impact on the integrity of the cell cytoskeleton, likely contributing to impair phagocytosis. Furthermore, it has been suggested that effectors of the T3SS are able to modulate the host's immune response. Here we present the first partial characterization of the immune response in rainbow trout (Oncorhynchus mykiss) infected with distinct strains of A. salmonicida either carrying (i) a fully functional T3SS or (ii) a functionally impaired T3SS or (iii) devoid of T3SS ("cured" strain). Infection with an A. salmonicida strain either carrying a fully functional or a secretion-impaired T3SS was associated with a strong and persistent immune suppression. However, the infection appeared to be fatal only in the presence of a fully functional T3SS. In contrast, the absence of T3SS was neither associated with immune suppression nor fish death. These findings suggest that the T3SS and T3SS-delivered effector molecules and toxins of A. salmonicida do not only impair the host cells' cytoskeleton thus damaging cell physiology and phagocytosis, but also heavily affect the transcription of critical immune mediators including the shut-down of important warning signals to recognize infection and induce immune defense.

Francisella Tularensis Clades B.FTN002-00 and B.13 Are Associated With Distinct Pathology in the European Brown Hare (Lepus europaeus).

Tularemia is a severe disease caused by Francisella tularensis This bacterium has a major pathogenic potential in countless animal species as well as in humans. Despite the relatively significant body of literature available on this microorganism, many questions are still open concerning its biological cycle in the environment, the pathology and pathogenesis of the disease, the possible routes of infection in animals, and the pathologic and ecological relevance of the distinct phylogenetic clusters of F. tularensis In order to address these questions, we have thoroughly characterized the pathology and microbiology of terminally ill European brown hares (Lepus europaeus) infected with F. tularensis subsp. holarctica, collected in Switzerland from 2012 to 2014. F tularensis isolates were typed by defining their phylogenetic clusters. We showed that the pathology associated with F. tularensis subsp. holarctica belonging to the clade B.FTNF002-00 is different from that previously reported to be associated with the clade B.13. In particular, strains of the clade B.FTNF002-00 were almost invariably associated with splenitis and hepatitis and not with the polyserositis affecting pleura, pericardium, and kidney reported in the literature for infections caused by the clade B.13. We describe findings suggesting that the ports of entry for the bacteria might be the respiratory and digestive routes.

Detection of Chlamydia pneumoniae in a collection of captive snakes and response to treatment with marbofloxacin.

In a collection of 58 snakes comprising predominantly Eurasian vipers in Switzerland, five snakes died unexpectedly during hibernation from 2009 to 2012. In one snake, organisms resembling chlamydiae were detected by immunohistochemistry in multiple histiocytic granulomas. Real-time quantitative PCR and microarray analysis were used to determine the presence of Chlamydia pneumoniae in tissue samples and cloacal/choanal swabs from snakes in the collection; 8/53 (15.1%) of the remaining snakes were positive. Although one infected snake had suppurative periglossitis, infection with C. pneumoniae did not appear to be associated with specific clinical signs in snakes. Of seven snakes treated with 5 mg/kg marbofloxacin IM once daily, five became PCR negative for C. pneumoniae following treatment, whereas one animal remained positive and one snake was lost to follow-up.

First detection of sarcoptic mange in free-ranging wild boar (Sus scrofa) in Switzerland.

In Switzerland sarcoptic mange is frequent in free-ranging wild carnivores but until recent years no cases had been recorded in wild ungulates. Since 2010, cases have been observed in wild boar in the cantons of Solothurn, Tessin and Thurgau. Here, we report the detection of mange-like skin lesions in wild boars by photo-trapping and the post-mortem findings in 6 culled animals presenting different stages of the disease. Potential sources of infection include mangy red foxes, outdoor domestic pigs and wild boars from surrounding countries. Disease spread in the wild boar population may become relevant not only for wildlife but also for domestic pig health in the future if piggeries' biosecurity is insufficient to prevent interactions with wild boar.

Characterisation of a new group of Francisella tularensis subsp. holarctica in Switzerland with altered antimicrobial susceptibilities, 1996 to 2013.

Molecular analysis of Francisella tularensis subsp. holarctica isolates from humans and animals revealed the presence of two subgroups belonging to the phylogenetic groups B.FTNF002-00 and B.13 in Switzerland. This finding suggests a broader spread of this group in Europe than previously reported. Until recently, only strains belonging to the Western European cluster (group B.FTNF002-00) had been isolated from tularaemia cases in Switzerland. The endemic strains belonging to group B.FTNF002-00 are sensitive to erythromycin, in contrast to the strains of the newly detected group B.13 that are resistant to this antibiotic. All the strains tested were susceptible to ciprofloxacin, streptomycin, gentamicin, nalidixic acid and chloramphenicol but showed reduced susceptibility to tetracycline when tested in a growth medium supplemented with divalent cations. The data show a previously undetected spread of group B.13 westwards in Europe, associated with changes in the antibiotic resistance profile relevant to treatment of tularaemia.

Fatal combined infection with canine distemper virus and orthopoxvirus in a group of Asian marmots (Marmota caudata).

A fatal combined infection with canine distemper virus (CDV) and orthopoxvirus (OPXV) in Asian marmots (Marmota caudata) is reported in this article. A total of 7 Asian marmots from a small zoological garden in Switzerland were found dead in hibernation during a routine check in the winter of 2011. The marmots died in February 2011. No clinical signs of disease were observed at any time. The viruses were detected in all individuals for which the tissues were available (n = 3). Detection of the viruses was performed by reverse transcription polymerase chain reaction. The most consistent gross lesion was a neck and thorax edema. A necrotizing pharyngitis and a multifocal necrotizing pneumonia were observed histologically. Numerous large intracytoplasmic eosinophilic inclusions were seen in the epithelial cells of the pharynx, of the airways, and in the skin keratinocytes. Brain lesions were limited to mild multifocal gliosis. Phylogenetic analysis revealed that the marmot CDV strain was closely related to the clusters of CDVs detected in Switzerland in wild carnivores during a local outbreak in 2002 and the 2009-2010 nationwide epidemic, suggesting a spillover of this virus from wildlife. The OPXV was most closely related to a strain of cowpoxvirus, a poxvirus species considered endemic in Europe. This is the first reported instance of CDV infection in a rodent species and of a combined CDV and OPXV infection.

Epidemic of salmonellosis in passerine birds in Switzerland with spillover to domestic cats.

A die-off of passerine birds, mostly Eurasian siskins (Carduelis spinus), occurred in multiple areas of Switzerland between February and March 2010. Several of the dead birds were submitted for full necropsy. Bacteriological examination was carried out on multiple tissues of each bird. At gross examination, common findings were light-tan nodules, 1 to 4 mm in diameter, scattered through the esophagus/crop. Histologically, a necroulcerative transmural esophagitis/ingluvitis was observed. Bacterial cultures yielded Salmonella enterica subsp. enterica serovar Typhimurium. At the same time, 2 pet clinics reported an unusual increase of domestic cats presented with fever, anorexia, occasionally dolent abdomen, and history of presumed consumption of passerine birds. Analysis of rectal swabs revealed the presence of S. Typhimurium in all tested cats. PFGE (pulsed field electrophoresis) analysis was performed to characterize and compare the bacterial isolates, and it revealed an indistinguishable pattern between all the avian and all but 1 of the feline isolates. Cloacal swabs collected from clinically healthy migrating Eurasian siskins (during autumn 2010) did not yield S. Typhimurium. The histological and bacteriological findings were consistent with a systemic infection caused by S. Typhimurium. Isolation of the same serovar from the dead birds and ill cats, along with the overlapping results of the PFGE analysis for all the animal species, confirmed a spillover from birds to cats through predation. The sudden increase of the number of siskins over the Swiss territory and their persistency during the whole winter of 2009-2010 is considered the most likely predisposing factor for the onset of the epidemic.

Emergence of canine distemper virus strains with modified molecular signature and enhanced neuronal tropism leading to high mortality in wild carnivores.

An ongoing canine distemper epidemic was first detected in Switzerland in the spring of 2009. Compared to previous local canine distemper outbreaks, it was characterized by unusually high morbidity and mortality, rapid spread over the country, and susceptibility of several wild carnivore species. Here, the authors describe the associated pathologic changes and phylogenetic and biological features of a multiple highly virulent canine distemper virus (CDV) strain detected in and/or isolated from red foxes (Vulpes vulpes), Eurasian badgers (Meles meles), stone (Martes foina) and pine (Martes martes) martens, from a Eurasian lynx (Lynx lynx), and a domestic dog. The main lesions included interstitial to bronchointerstitial pneumonia and meningopolioencephalitis, whereas demyelination--the classic presentation of CDV infection--was observed in few cases only. In the brain lesions, viral inclusions were mainly in the nuclei of the neurons. Some significant differences in brain and lung lesions were observed between foxes and mustelids. Swiss CDV isolates shared together with a Hungarian CDV strain detected in 2004. In vitro analysis of the hemagglutinin protein from one of the Swiss CDV strains revealed functional and structural differences from that of the reference strain A75/17, with the Swiss strain showing increased surface expression and binding efficiency to the signaling lymphocyte activation molecule (SLAM). These features might be part of a novel molecular signature, which might have contributed to an increase in virus pathogenicity, partially explaining the high morbidity and mortality, the rapid spread, and the large host spectrum observed in this outbreak.

Glomerulonephropathy in aged captive Key Largo woodrats (Neotoma floridana smalli).

A retrospective review of mortality records of Key Largo woodrats (Neotoma floridana smalli) in a captive breeding program revealed chronic renal disease in 5 of 6 woodrats older than 4 years of age. Two of the 5 woodrats with chronic renal disease also had clinical evidence of diabetes mellitus. Kidneys from all 5 woodrats were examined via light microscopy, histochemical staining, immunohistochemical staining, and transmission electron microscopy. The dietary histories of the affected animals were examined as well. The most striking histopathologic abnormality in the affected kidneys was the presence of large protein casts within cortical and medullary tubules in combination with lesions of membranous glomerulopathy and glomerulosclerosis. Transmission electron microscopy revealed thickening and undulation of the tubular and glomerular mesangial basement membranes with the variable presence of electron-dense deposits within the capillary endothelial basement membrane. Patchy glomerular immunoreactivity for IgG was noted in 2 cases, but IgA and IgM immunoreactivity were not present. The pathologic changes in the kidneys of the Key Largo woodrats mirrored many of the features of chronic progressive nephropathy commonly diagnosed in laboratory rats. Woodrats in the captive population were fed an ad libitum high-protein diet similar to diets that have been shown in laboratory rats to exacerbate the development and progression of chronic progressive nephropathy. It is concluded that Key Largo woodrats develop glomerulonephropathy with features similar to chronic progressive nephropathy described in laboratory rats. Age, concomitant disease, and dietary factors may contribute to the development and severity of this potentially age-limiting disease in Key Largo woodrats.

Experimental transmission of a herpesvirus in Greek tortoises (Testudo graeca).

An experimental transmission study aimed at fulfilling Koch's postulates for a herpesvirus-associated stomatitis-rhinitis in Mediterranean tortoises is presented. Clinical, pathologic, serologic, and molecular studies were performed linking tortoise herpesvirus with the pathogenesis of stomatitis-rhinitis. Four adult Greek tortoises received either intranasally or intramuscularly two tortoise herpesvirus isolates by primary experimental infection and secondary challenge 11 months later. After the primary experimental infection and the secondary challenge, clinical signs of illness developed, which included conjunctivitis, diphtheritic oral plaques, and oral discharge. At 4 weeks after the secondary challenge, all tortoises were humanely euthanatized and evaluated. Although neutralizing antibodies developed after the primary experimental infection, they apparently did not prevent the later development of recurrent clinical signs. Polymerase chain reaction (PCR) and reverse transcription-PCR analyses allowed sensitive characterization of the systemic distribution of the herpesvirus DNA sequences and their presence in the cranial nerves and brains of the infected tortoises. Despite the failure to recover the herpesviruses used in the transmission study, the findings support the premise that tortoise herpes-virus is a primary pathogen of Greek tortoises.

Application of immunoperoxidase-based techniques to detect herpesvirus infection in tortoises.

Indirect (IIP) and direct (DIP) immunoperoxidase assays were developed for the serological and histological diagnoses of herpesvirus infection in tortoises, respectively. A mouse monoclonal antibody (MAb HL1546), specific for the heavy chain of tortoise IgY, was used as the secondary antibody in the IIP assay. Rabbit polyclonal antisera raised against 2 sucrose gradient-purified tortoise herpesvirus isolates (HV4295/7R/95 and HV1976) were used as primary antibodies for the detection of herpesvirus antigen either in infected cell cultures or in formalin-fixed, paraffin-embedded tissues. The IIP and DIP assays could detect either the presence of anti-herpesvirus antibody in the plasma of exposed tortoises or the presence of herpesvirus antigen in infected tissues, respectively. Although the IIP test complements the enzyme-linked immunosorbent assay and the serum neutralization test already available for measuring herpesvirus-specific antibody in tortoises, the DIP test is useful for the histological diagnosis of herpesvirus infection in tortoises.

Enzyme-linked immunosorbent assay for detecting herpesvirus exposure in Mediterranean tortoises (spur-thighed tortoise [Testudo graeca] and Hermann's tortoise [Testudo hermanni]).

An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of antibodies to a herpesvirus associated with an upper respiratory tract disease in Mediterranean tortoises [spur-thighed tortoise (Testudo graeca) and Hermann's tortoise (Testudo hermanni)]. This serodiagnostic test was validated through a hyperimmunization study. The mean of the A(405) readings of the plasma samples collected at time zero of the hyperimmunization study plus three times the standard deviation was used as the cutoff for seropositivity in tortoises. ELISA results were compared to serum neutralization (SN) values for the same samples by using the McNemar test. The results obtained by SN and ELISA were not significantly different (P > 0.05). This new ELISA could be used as an important diagnostic tool for screening wild populations and private and zoo collections of Mediterranean tortoises.

Diseases of the respiratory tract of chelonians.

Diseases of the respiratory tract commonly occur in captive chelonians, and several diseases also have occurred in wild chelonians. Infectious causes include viruses, bacteria, fungi, and parasites. Herpesviruses have surfaced as important pathogens of the oral cavity and respiratory tract in Hermann's tortoise (Testudo hermanii), spur-thighed tortoise (Testudo graeca), and other tortoises in Europe and the United States. Herpesvirus-associated respiratory diseases also have been reported in the green turtle, Chelonia mydas, in mariculture in the Cayman Islands. Of diseases caused by bacteria, an upper respiratory tract disease caused by Mycoplasma sp has been reported in free-hanging and captive gopher tortoises in the southeastern United States and in desert tortoises in the Mojave Desert of the southwestern United States. Mycotic pulmonary disease is commonly reported in captive chelonians, especially in those maintained at suboptimal temperatures. An intranuclear coccidia has been seen in several species of captive tortoises in the United States, and, in one case, a severe proliferative pneumonia was associated with organisms in the lung. The most common noninfectious cause of respiratory disease in chelonians results from trauma to the carapace. Although pulmonary fibromas commonly occur in green turtles with fibropapillomatosis, for the most part, tumors of the respiratory tract are uncommon in chelonians.