Parasitological examination results of zoo animals in Germany between 2012 and 2022.

Parasitic infections in zoo animals are a critical concern for both animal health and management. The aim of this study was to assess the occurrence of endo- and ectoparasites among zoo animals in Germany. A retrospective analysis of the submitted samples of a diverse range of zoo animals (5768) from a ten-year period (2012-2022) was conducted. Overall, 31.1% of those samples tested positive for at least one parasite. In the examined samples, helminths (28.4%) were found more often than protozoans (10.3%) or ectoparasites (0.8%). Among the various animal groups the following parasites were found most commonly: Artiodactyla: Coccidia (34.6%), Strongylida (23.4%); Perissodactyla: Strongylida (19.3%), Ascaridida (12.0%); Carnivora: Ascaridida (16.6%), Coccidia (8.1%); Rodentia: Oxyurida (18.2%), Coccidia (10.5%); Marsupialia: Coccidia (9.4%), Oxyurida (5.9%); Primates: Trichuris spp. (9.7%), Oxyurida (2.2%); Aves: Capillaria (7.8%), Ascaridida (7.6%); Reptilia, Amphibia, Insecta: Oxyurida (18.7%); Pisces: Ciliates (6.2%). Furthermore, potentially zoonotic parasites were identified, including Toxoplasma gondii (0.1%), Cryptosporidium sp. (0.1%). By examining the occurrence of specific parasites, these findings demonstrate the importance of parasites in the context of zoo animal health. They also highlight the need for effective strategies to control parasite burden to improve the overall welfare of zoo animals.

Interplay between Eimeria acervulina and Cryptosporidium parvum during In Vitro Infection of a Chicken Macrophage Cell Line (HD11).

BACKGROUND: Eimeria acervulina is a frequent intestinal pathogen of chickens, causing economic impact on the poultry industry. Cryptosporidium parvum is a neglected parasite in chickens. However, because of its zoonotic potential, poultry cryptosporidiosis may pose a risk to public health. Little is known about the parasite-host interactions during coinfection with both parasites. In this study, we investigated the possible interactions during in vitro coinfection of E. acervulina and C. parvum in a chicken macrophage cell line (HD11). METHODS: HD11 cells were inoculated with E. acervulina and C. parvum sporozoites and incubated 2, 6, 12, 24, and 48 h post infection (hpi). Mono-infections for each parasite were also investigated. Real-time PCR was used to quantify parasite replication. Additionally, macrophage mRNA expression levels of IFN-γ, TNF-α, iNOS, and IL-10 were measured. RESULTS: For both parasites, multiplication was, in most groups, lower in the coinfection group (COIG) compared with mono-infections. However, at 6 hpi, the number of C. parvum copies was higher in co-infections. Intracellular replication started to decrease from 12 hpi onward, and it was almost undetectable by 48 hpi in all groups. Infections resulted in low expression of all cytokines, except at 48 hpi. CONCLUSIONS: Infection of avian macrophages with both E. acervulina and C. parvum seemed to hinder intracellular replication for both parasites in comparison to mono-infection. A clear reduction in intracellular parasites from 12 hpi onward details the important role potentially played by macrophages in host control of these parasites.

Morphological and Molecular Identification of Physaloptera alata (Nematoda: Spirurida) in a Booted Eagle (Aquila pennata) from Portugal.

Physaloptera spp. are parasitic nematodes that infect the gastrointestinal tracts of many carnivores and omnivores. Although they are distributed worldwide, Physaloptera spp. have not been studied in raptors in Portugal. In this study, we report Physaloptera alata in a booted eagle (Aquila pennata) in Portugal. Adult nematodes were discovered in the gizzard of a young booted eagle, and morphological features were consistent with those of the genus Physaloptera. DNA was extracted and a PCR assay performed to amplify a region of the 18S small subunit of the ribosomal RNA gene and the cytochrome c oxidase subunit I gene. The resulting PCR products were Sanger-sequenced, and comparison with the available sequences in the GenBank database confirmed the initial morphological classification as Physaloptera sp. Phylogenetic analysis clustered the sequence within the Physaloptera group. The presence of this parasite in raptors from Portugal is of particular importance to wildlife rehabilitation centers, disease ecologists, and wildlife professionals. Furthermore, we produced a new genetic sequence and have added it to the GenBank database of parasites in birds of prey.

Diplotriaena obtusa infection in an Eurasian blue tit (Cyanistes caeruleus) in Germany. Pathology and phylogenetic analysis.

Diplotriaena obtusa is a nematode found in air sacs of a wide number of birds, including Passerine species. During a period of increased mortality of blue tits (Cyanistes caeruleus) in Germany, we collected adult nematode worms from the air sacs of a deceased male blue tit. The nematodes showed morphological features consistent with Diplotriaena ssp. A polymerase chain reaction (PCR) assay targeting the small subunit (SSU) 18S rRNA gene identified the parasite species as Diplotriaena obtusa. Phylogenetic analysis confirmed species identification. Further examinations against infectious pathogens like Suttonella ornithocola, Salmonella spp., Pasteurella spp., Chlamydia spp., Influenza A virus, Usutu virus and West Nile virus were negative. This is the first report of D. obtusa in a blue tit from Germany.

First detection of Echinococcus granulosus sensu stricto (G1) in dogs in central Sudan.

Eighty-four stray dogs shot as a part of a governmental rabies control program in two neighboring towns of central Sudan were examined for the presence of Echinococcus spp. and other intestinal helminths. Echinococcus worms were identified to species level by PCR and gene sequencing. For comparative reasons, rectal content of the necropsied dogs was examined for helminth eggs and subjected to copro-PCR for Echinococcus. At necropsy, 51.2% (43/84) of the dogs harbored Echinococcus canadensis (G6/7) worms with worm burdens ranging from 22,000 to 80,000. Dipylidiun caninum was found in 53.6% of the dogs. At coproscopy, taeniid eggs were found in 37 of the 43 dogs which were positive for Echinococcus at necropsy, but none in the 41 necropsy-negative dogs. In addition, 58% of the rectal samples contained eggs of Toxocara spp., 34.5% eggs of Trichuris spp. (34.5%), and 26% eggs of Ancylostoma caninum. Copro-PCR gave positive results for E. canadensis with 97.5% (39/40) of nonhibiting samples from the necropsy positive dogs; the one remaining dog tested positive for E. granulosus sensu stricto (G1), whose partial cox1 and nad1 sequences showed a 100% identity with various reference sequences of the G1 genotype. 100% of 38 non-inhibited samples taken from the necropsy-negative dogs were also negative in copro-PCR. This is the first study which combines prevalence and genetic identification of Echinococcus spp. in dogs of Sudan. Together with a recent report from cattle, it confirms the autochthonous presence, at low level, of E. granulosus sensu stricto in Central Sudan.

Diagnostics and epidemiology of alveolar echinococcosis in slaughtered pigs from large-scale husbandries in Germany.

By means of the official meat inspection of domestic pigs, exceptionally high proportions of livers affected by encapsulated nodules containing whitish to light yellow, viscous to pasty material ("microabscesses") were detected. The swine had been raised on four different farms, being located in distinct regions of Germany (Brandenburg, Thuringia, Upper Franconia). Macroscopical and histological examination of 77 samples of livers revealed granulomatous to necrotizing hepatitis with attendance of numerous eosinophils. In 61 % (n = 47) of the lesions, eosinophilic, band-like acellular structures resembling the laminated layer of Echinococcus sp. were visible. Moreover, representative samples (n = 11) showed a positive reaction of these structures with Periodic acid-Schiff. Altogether, the findings were consistent with alveolar echinococcosis. Echinococcus multilocularis DNA could be demonstrated in selected samples (n = 7) by polymerase chain reaction. Epidemiological considerations suggest contamination of the forage with fox tapeworm eggs to be the most likely source of infection on two of the farms, as some of the fodder had been stored in the open, being amenable to infected definitive hosts. On the two other farms, mainly straw litter has to be taken into account regarding the transmission route, since carnivores excreting eggs of E. multilocularis could have gained access to the straw storage. The presented cases show that adequate mechanisms of meat inspection may provide important data for the purposes of surveillance and risk assessment of human alveolar echinococcosis.

Quantitative comparison of different purification and detection methods for Cryptosporidium parvum oocysts.

UNLABELLED: Cryptosporidium parvum (C. parvum) is the causal agent of cryptosporidiosis in many animals, mainly cattle, and possesses a high zoonotic potential. It occurs worldwide and ubiquitously. Detection of C. parvum is mainly performed directly but purification of the oocysts is useful to increase sensitivity and to obtain oocyst material for further use. The study was designed to compare (a) three different direct diagnostic methods, namely modified Ziehl-Neelsen staining, carbol fuchsin staining and conventional PCR, and (b) three routine oocyst purification methods, in particular flotation with saturated sodium chloride solution, Sheather's sucrose solution and a Percoll(®) gradient. During comparison of purification methods, special regard was paid to the ability to separate morphologically intact oocysts from the morphologically degenerated fraction or viable from non-viable oocysts, respectively. RESULTS: (a) DIAGNOSTIC METHODS: Most effective in C. parvum oocysts detection in calf faeces was PCR; carbol fuchsin and modified Ziehl-Neelsen stainings achieved comparable results. (b) Purification methods: Oocyst flotation using sodium chloride solution showed to be superior to Percoll(®) gradient centrifugation and sugar flotation in terms of purification quality, recovery efficacy (yield) and reduction of the proportion of degenerated or non-viable oocysts.

Echinococcus multilocularis infections in domestic dogs and cats from Germany and other European countries.

A cross-sectional survey was conducted to estimate the prevalence of Echinococcus multilocularis and E. granulosus infections in domestic dogs and cats from Germany and other European countries. Faecal samples of 21,588 dogs and 10,650 cats routinely submitted to a private veterinary laboratory between June 2004 and June 2005 were examined using the ZnSO(4)-NaCl flotation method. Taeniid eggs were detected in 54 (0.25%) and 37 (0.34%) of the canine and feline faecal samples, respectively. Taeniid eggs were separated and subjected to a DNA preparation and a modified two-step PCR for the detection of Echinococcus spp. based on mitochondrial 12S rRNA genes. PCR products from Echinococcus-negative but cestode-positive reactions were cloned and sequenced to determine the Taenia species. E. multilocularis DNA was specifically amplified in 43 (0.24%) and 25 (0.23%) of the samples from dogs and cats, respectively. E. granulosus DNA was not detected in any sample, while, E. multilocularis-positive samples were detected in dogs from Germany only, those of cats originated from Germany, Denmark and The Netherlands. The prevalence of E. multilocularis egg-positive canine samples was significantly higher in southern (0.35%) than in northern Germany (0.13%). In contrast, no significant regional difference was observed in cats from Germany. Taeniid eggs from Echinococcus-negative samples and from a few samples with macroscopically detected Taenia sp. proglottids were identified as eggs of T. crassiceps (n=8), T. martis, T. serialis, T. polyacantha, T. taeniaeformis and T. pisiformis in dogs (n=1 of each) and T. taeniaeformis (n=11) in cats. The spectrum of cestodes detected in domestic dogs and cats indicate the consumption of small rodents as infection source. The high proportion of E. multilocularis-positive samples, suggest domestic dogs and cats as a possible source of E. multilocularis infection for humans.