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Contrary to humans, candidiasis is a rare infection in animals. However, in reptiles, candidiasis can cause gastrointestinal, cutaneous, or rarely systemic infections in stressed animals. The infections due to

Simple Summary The European hedgehog is a very common animal in Europe. This animal usually lives in woods but often it is possible to find it in gardens and parks in urban areas. Few studies describe parasites infecting the European hedgehog in Italy. In the present study, endoparasite infections and associated lesions were investigated in hedgehogs from two different regions of central Italy (Latium and Tuscany), where no investigations had been performed before. Several helminth and protozoan species were identified in the intestine and respiratory tract of the animals analyzed. The presence of the respiratory worm Crenosoma striatum was found to be associated with bronchial lesions. This study contributed to the knowledge of parasitic infections of the European hedgehog, the most frequently hospitalized animal in wildlife rescue centers. Abstract The European hedgehog is a synanthropic mammal, widely distributed in Europe. This species usually inhabits the edges of deciduous or mixed woods, but it is also very common in private gardens and public parks. Despite its popularity and frequency of contacts both with humans and with wild and domestic animals, few studies have examined the endoparasitic fauna of the hedgehog in Italy. In the present study, endoparasites of naturally deceased hedgehogs (n = 40) from central Italy (Latium and Tuscany regions) were investigated, along with concurrent gross and histopathological lesions. The most prevalent identified endoparasites were Crenosoma striatum (45%), Capillaria erinacei (42.5%) and Brachylaemus erinacei (22.5%), in accordance with previous reports from hedgehogs in southern Italy. In few subjects, Physaloptera clausa, Acanthocephalans and Cystoisospora rastegaeivae coccidia were also identified. The infection by the lungworm C. striatum was found to be significantly associated (p < 0.01) with bronchial hyperplasia and peribronchiolitis upon histopathological examination. Awareness of the most common parasitic infections in the hedgehog and of their effects on the health of these animals is extremely important, especially in wildlife rescue centers, where European hedgehog represents the most frequently hospitalized mammal species.

The genus Pearsonema, in the nematode family Capillariidae, includes several species that parasitize the urinary bladders of wild and domestic carnivores. The infection has been reported worldwide from several wildlife species, including canids, mustelids, and felids, but the pathological aspects have seldom been investigated. In order to assess the presence and severity of the lesions in Pearsonema-infected wildlife, we performed a parasitological and pathological examination of urinary bladders from 72 animals, belonging to the families Canidae (red fox Vulpes vulpes, n = 28, and wolf Canis lupus, n = 29) and Mustelidae (beech marten Martes foina, n = 3, pine marten Martes martes, n = 2, and European badger Meles meles, n = 10). A greater prevalence of infection for canids (64.91%, 95% confidence interval (95% CI), 52.52–77.30%) than for mustelids (13.33%) (p &lt, 0.001) was recorded. The prevalence of infection in red foxes was 75.0% (95% CI, 58.96–91.04%), in accordance with other reports from European countries, supporting the role of this species as a reservoir for infection. Eosinophilic cystitis was observed in 34 out of the 72 examined animals (47.22%). The influence of Pearsonema sp. infection on the occurrence of eosinophilic cystitis was statistically significant in wolves (p &lt, 0.01), which were also affected by more severe histological lesions compared to foxes.

With the aim to identify the Eimeria species responsible for coccidiosis in 50 deceased red-legged partridges (Alectoris rufa), individual faecal samples were collected, dissolved in 2.5% K2Cr2O7 solution and maintained at room temperature to allow sporulation of the oocysts. Morphology and dimensions of sporulated oocysts were microscopically evaluated. To assess Eimeria intestinal localisation, faecal samples and scrapings taken from the different intestinal segments of each deceased animal were examined by fresh smears and flotation test, while the intestines were examined for gross lesions, then fixed in 10% formalin and processed for histopathological analysis. From scrapings and morphological analysis, Eimeria kofoidi and Eimeria legionensis were identified in the small intestine and in the caecum and colon, respectively. Histopathological analysis confirmed the presence of two distinct Eimeria species. In particular, E. kofoidi macrogamonts were found in epithelial cells of jejunum and ileum, between the basal lamina and the nucleus of the infected intestinal cells. This latter was flattened and displaced above. E. legionensis macrogamonts were instead found localised between the nucleus and the luminal surface of the infected caeca and colonic cells and these macrogamonts were larger than those of E. kofoidi. Chronic enteritis and severe displacement of the deep crypts of the small intestine, large areas of caeca and colonic epithelial necrosis associated to thickened wall and mononuclear cells infiltration diffused in a transmural manner, were the main histopathological lesions.

Parasite monitoring is considered a necessary step for cetacean management and conservation. Between February 2013 and July 2015, 26 dolphins (15 Stenella coeruleoalba, 10 Tursiops truncatus, and one Grampus griseus) stranded along the Tuscan coastline of the protected marine area &ldquo, Pelagos Sanctuary&rdquo, were examined. Organs, tissues, and faecal and blood samples taken from all animals were analysed by parasitological, immunological, and molecular techniques. Twenty-one out of 26 dolphins (80.77%) tested positive for at least one parasite species, and 13/15 (86.7%) S. coeruleoalba, 7/10 (70%) T. truncatus, and the single G. griseus were found positive. Identified parasites included the nematodes Skrjabinalius guevarai (7.69%, 2/26), Halocercus lagenorhynchi (3.85%, 1/26), Halocercus delphini (7.69%, 2/26), Stenurus ovatus (7.69%, 2/26), Crassicauda spp. (7.69%, 2/26), the trematodes Pholeter gastrophilus (26.92%, 7/26), Campula palliata (3.85%, 1/26), the cestodes Phyllobothrium delphini (42.31%, 11/26), Monorygma grimaldii (23.08%, 6/26), Tetrabothrium forsteri (7.69%, 2/26), Strobilocephalus triangularis (7.69%, 2/26), and the acanthocephalan Bolbosoma vasculosum (7.69%, 2/26). Moreover, 6/26 (23%) animals scored positive to Toxoplasma gondii at serology, but PCR confirmed the infection (T. gondii Type II genotype) in a single animal. In examined dolphins, obtained results showed a high prevalence of endoparasites, which included species considered as a cause of severe debilitation or death.

Tattoo skin disease (TSD) is a poxviral disease typical of cetaceans. Two juvenile and well-preserved male striped dolphins (Stenella coeruleoalba), found stranded along the Tuscany and Latium coasts of Italy in 2015 and 2016, respectively, showed typical skin lesions ascribable to TSD. Histological, ultrastructural and biomolecular investigations confirmed a poxviral aetiology for the aforementioned skin lesions. To our knowledge, this should be the first report of TSD in cetaceans stranded along the Italian coastline. As organochlorines like PCBs and DDTs are known to be highly immunotoxic, the tissue loads of these contaminants were evaluated, in order to increase our knowledge on their potential role as well as on the relationships between the level of exposure to these pollutants and poxviral infection's occurrence.

Ciliates have been reported as pathogens of many species of economically important bivalves. Mussel protozoan X (MPX), is an uncharacterised intracellular ciliate of mussels and has been widely reported in Mytilus spp. around the world. In order to characterise this ciliate, Mytilus edulis samples were collected from a site on the West coast of Scotland, and four different fixatives for histological examination were tested. Fresh preparations of mussel digestive glands were also examined by laser scanning confocal microscopy. Intracellular ciliates were prepared by laser capture microdissection and partial sequences of small subunit ribosomal RNA gene and of large subunit ribosomal RNA gene were generated, using Phyllopharyngea primers. Methacarn solution proved to be the best fixative for both histological and molecular characterisation. The morphological and molecular investigations confirmed that this ciliate belongs to the class Phyllopharyngea, order Rhynchodida. However, this organism does not belong to any known family, genus or species, therefore, a new description is necessary, following further morphological analyses. Most mussel samples containing MPX displayed mild to moderate infections, with no signs of necrosis or haemocytic response, although a single sample displayed a severe infection (∼103 ciliates per section). The localisation of this ciliate in tissues other than the digestive gland, the presence of necrosis in infected tissue of the most severely infected mussel and the binary fission of this ciliate have been observed here for the first time. We also report the first observation of the live ciliate isolated from tissue. Although MPX remains of unknown significance to the mussel industry, tools and protocols described here will be useful in further characterising these and other ciliates (subclass Rhynchodia) known as pathogens for bivalves.

The in vitro anthelmintic activity on sheep gastrointestinal strongyle (GIS) eggs and larvae of 0.5% aloin and 0.1% aloe-emodin was investigated. From fresh faecal samples collected by ewes naturally infected by Haemonchus, Trichostrongylus and Teladorsagia nematodes, GIS eggs were isolated and cultivated in Petri dishes (100 eggs/dish). For the in vitro evaluation of the anthelmintic activity of tested compounds, the Egg hatch test (EHT), the Larval development test (LDT) and the Larval mortality/paralysis test (LMT) were used. In each assay, the activity of tested compounds was compared to untreated and treated (0.1% thiabendazole, TBZ) controls. Six repetitions were made through the experiment. Obtained data were statistically elaborated using the X2 test. In EHT, 0.5% aloin gave highly significantly different (P1 to L3, showing no significant differences (P

In September 2014, seven sperm whales were stranded along Italy’s Adriatic coastline. Postmortem investigations on 3 female adult whales and 1 male fetus carried by the largest female revealed molecular and immunohistochemical evidence of dolphin morbillivirus infection. A possible role of the virus in the stranding event was considered.

[EN] The dynamic change in faecal Eimeria oocyst excretion was evaluated in 10 naturally infected female rabbits, starting from their weaning at 33 d of age until about 1 mo after their second parturition. Faecal samples collected from examined animals were quali-quantitatively analysed to evaluate presence and number of Eimeria oocysts. In addition, isolated Eimeria oocysts were identified at the species level following sporulation. Animals were found to be infected by Eimeria perforans, Eimeria exigua and Eimeria magna and shed Eimeria oocysts after weaning and after parturition. In particular, at 33 d of age all female rabbits examined were negative, while the discharge of Eimeria oocysts started at 39th day of age and peaked between 46th and 53rd day of age. From 81-109 d of age until the first parturition and from 25 d of age of the litters born at the first parturition to the second parturition, all animals resulted negative. After parturition, Eimeria oocyst output occurred from 6th to 12th day after the first parturition and from 7th to 13th day after the second parturition, while a second period of oocyst excretion was observed from 18th to 24th day after both parturitions. These findings may indicate the existence of a relationship between the periparturient phase and Eimeria oocyst output and suggest an important role of the mothers in transmission of the infection to their litters., The authors thank the Italian Ministry of University (MIUR) for financing this study

Photobacterium damselae subsp. damselae (PDD) is a known pathogen of fish, humans and marine mammals. In this study, a Multilocus Sequence Typing (MLST) scheme based on six housekeeping genes (glp, gyrB, metG, pnt, pyrC, and toxR) was developed to better understand the PDD population structure and used to type 73 PDD isolates from cetaceans, mainly striped dolphins (Stenella coeruleoalba) involved in mortality episodes, and from a few marine chelonians. Five reference ATCC strains were also included in the study. Typing allowed the discrimination of groups of PDD strains isolated from different host species, at different times and from different geographic areas, suggesting that a clonal PDD group may have spread in the Tyrrhenian sea at the time of an Unusual Mortality Event (UME) among cetaceans, mainly striped dolphins, occurred in early 2013 along the Italian western coasts.

In captive reptiles, parasites may represent a serious health and management problem for both the reptiles and their human caretakers (zoonotic parasites). In the present study, the prevalence of ecto- and endoparasitic infections and the clinical signs associated with these parasites were determined in pet reptiles living in Italy. To accomplish these aims, fecal samples collected from 120 captive reptiles representing 27 different species, including 63 household pet and 57 pet store reptiles, were analyzed macroscopically and microscopically for the isolation and identification of endoparasites. In addition, the reptiles, along with their terrariums and water containers, were also examined for ectoparasitic arthropods. Recovered arthropods were preserved in 80% ethanol and evaluated microscopically. Overall, 72.5% of the examined reptiles were found to be positive for parasites, with prevalence rates of 45% and of 27.5% for endo- and ectoparasites, respectively. Pet shop animals were more likel...

By using the egg hatch test (EHT), the larval development test (LDT) and the larval mortality/paralysis test (LMT), the in vitro anthelmintic activity on sheep gastrointestinal strongyles (GIS) of four plant-derived pure compounds, mangiferin (at 0.25%, 0.125% and 0.0625%), rutin (at 1%, 0.75%, 0.5%), quercetin (at 1%), and &beta, sitosterol (at 1%, 0.75%, 0.5%), was investigated. For comparison, untreated and treated (0.1% thiabendazole, 0.1% TBZ) controls were used. Six repetitions were made throughout the experiment. Data were statistically elaborated using the &chi, 2 test. The concentration able to inhibit the development of the 50% of L1s to L3s and causing the mortality of the 50% of L3s (EC50) was also calculated. L3s recovered from untreated Petri dishes were identified at the genus level. In EHT, all tested compounds at all concentrations significantly (p &lt, 0.01) inhibited the hatch of the eggs when compared to the untreated controls, but none of them was as effective as 0.1% TBZ. In LDT, rutin (at 1%, 0.75% and 0.5%), mangiferin (at 0.25% and 0.125%), &beta, sitosterol (at 1%) and 0.1% TBZ completely prevented the larval development from L1 to L3 in respect to the untreated controls (p &lt, 0.01). In LMT, all tested compounds significantly (p &lt, 0.01) increased the death of L3s compared to the untreated controls, except for &beta, sitosterol at 0.5%. However, only rutin at all concentrations and 0.25% and 0.125% mangiferin were as effective as 0.1% TBZ. Haemonchus, Trichostrongylus, Chabertia and Teladorsagia/Ostertagia GIS genera, were identified.

Prevalence and risk factors of Leishmania infantum, Dirofilaria spp. and other potentially zoonotic or canine-specific endoparasite infections were assessed in 639 kennel dogs from central Italy. To this end, individual blood and fecal samples were examined using parasitological, immunological and molecular techniques. The presence of compatible clinical pictures, as well as age and gender were considered as putative risks factors. To evaluate risk factors, multivariable analysis with logistic regression and univariable analysis with a Chi square test and a Fischer’s exact test were performed. Overall, 52.6% of dogs (95% CI 48.6-56.5) were found positive, while 39.6% of dogs (95% CI 35.8-43.5) were infected by potentially zoonotic species. Leishmania infantum and Dirofilaria repens showed prevalences of 2.5% (95% CI 1.5-4.1) and 2.8% (95% CI 1.7-4.5), respectively. The prevalence of cardiorespiratory parasites was 7.8% (95% CI 5.9-10.3) and included the species Angiostrongylus vasorum, Eucoleus aerophilus, Eucoleus boehmi and D. immitis; the latter showed a prevalence of 0.2% (95% CI 0.001-1). Intestinal parasites were significantly prevalent (38.8%, 95% CI 35-42.7) and they consisted mainly of species of major zoonotic concern, including ancylostomatids, Toxocara canis, Giardia duodenalis, Dipylidium caninum, Taeniidae, Strongyloides stercoralis and Cryptosporidium parvum. Endoparasites were significantly prevalent in clinically suspected dogs. Leishmania infantum and cardiorespiratory nematodes were prevalent in older dogs, while intestinal parasites were prevalent in younger dogs. Results show high dog and public health risks in kennels in central Italy, and suggest the need for more effective control measures.

In this study, some in vitro trials were carried out to evaluate the association between Scopulariopsis spp. fungi with Psoroptes cuniculi (Acari: Psoroptidae) and their potential pathogenicity to this mite species. After cultivation on Sabouraud dextrose agar with chloroamphenicol at 26 degrees C for 20 days and macro- and microscopical examinations, from P. cuniculi mites taken from some infested rabbits fungi belonging to the genus Scopulariopsis were isolated. Investigations were carried out to evaluate in vitro the potential pathogenic role of Scopulariopsis to P. cuniculi; to this aim, Scopulariopsis brevicaulis isolated from a cat was tested. In several culture media, the dose-dependent P. cuniculi mortality with different concentrations of S. brevicaulis and the ability of S. brevicaulis to penetrate inside the body of infected mites were evaluated. Results obtained demonstrated that, in the rabbit, Scopulariopsis fungi can be associated with P. cuniculi, and that S. brevicaulis can be an entomopathogen for P. cuniculi in a dose-dependent manner. A more rapid mortality of the mites and a quicker S. brevicaulis growth in plates containing rabbit serum with respect to all other media used were also found. S. brevicaulis demonstrated the ability to invade the body of P. cuniculi.

The acaricidal activity of Eugenia caryophyllata essential oil was evaluated in vitro and in vivo on Psoroptes cuniculi, a mange mite. In vitro, different concentrations of the oil were tested and the observed mites mortality was compared with that observed in untreated and treated (Acacerulen R) controls. In vivo, six P. cuniculi infected rabbits were topically treated with the oil diluted at 2.5% and compared with untreated and treated control groups of six rabbits each. In vitro, up to the concentration of 0.10% the oil gave highly significant (P

During 2014, six loggerhead turtles, Caretta caretta and one green turtle, Chelonia mydas, found stranded on the Tuscany coast of Italy, were examined for the presence of specific bacterial and viral agents, along with their role as carriers of fish and human pathogens. Thirteen different species of bacteria, 10 Gram negative and 3 Gram positive, were identified. Among them, two strains of Vibrio parahaemolyticus and one strain of Lactococcus garviae were recovered and confirmed by specific PCR protocols. No trh and tdh genes were detected in V. parahaemolyticus. The first isolation of L. garviae and the first detection of Betanodavirus in sea turtles indicate the possibility for sea turtles to act as carriers of fish pathogens. Furthermore, the isolation of two strains of V. parahaemolyticus highlights the possible role of these animals in human pathogens' diffusion.

An unusual mortality event involving cetaceans, mainly striped dolphins Stenella coeruleoalba (Meyen, 1833), occurred along the Tyrrhenian Sea coast of Italy during the first 3 mo of 2013. Based on post-mortem analyses carried out according to body condition on 66 dolphins (54% of stranded animals), several hypotheses to explain the causes of this mortality event were proposed. Although no definitive conclusions can be drawn, dolphin morbillivirus was deemed the most likely cause, although other infectious agents (including Photobacterium damselae damselae and herpesvirus) or environmental factors may also have contributed to this recent mortality event.

Parakeets and parrots can be infected by several parasitic species that may be either primary or opportunistic pathogens and may be responsible for mild or severe clinical forms of disease (Greiner and Ritchie 1994, Doneley 2009). Some psittacine parasitic species are host specific, while others can infect a wide range of animals—including humans (Tsai and others 1992, Fayer 2010). The main aim of this study was the evaluation of the prevalence of ectoparasites and endoparasites in captive psittacine birds living in Italy. In the period 2004–2007, 573 randomly selected captive psittacine birds of 47 different species living in Italy were examined. In this prospective survey all animals, including 450 farmed and 123 pet psittacines, were screened for endoparasites, while 196 farmed and 25 pet birds were examined also for ectoparasites. For endoparasites, fresh individual faecal samples were analysed by flotation test, a commercial immunoassay (RIDAQUICK Cryptosporidium / Giardia Combi cassettes, R-Biopharm, Italy) and by Giemsa and modified Ziehl Neelsen stained faecal smears. In addition, a deceased Platycercus elegans and a deceased Ara chloroptera were necropsied and examined for gross lesions and parasites. Isolated nematodes were identified at the species level (Levine 1968, Kajerova and others 2004a, b). Species of collected ectoparasites were also identified (Krantz 1978, Atyeo and Gaud 1987, Mironov and Galloway 2002, Schmaschke and others 2002, Sychra and others 2007, Di Palma and others …

The development of Eucoleus bohmi eggs was evaluated in vitro in order to acquire information on the life cycle of this neglected respiratory nematode affecting canids. Fecal cultures were prepared using fecal samples from a positive dog and maintained at different conditions of temperature (20 ± 1 and 5 ± 1 °C) and relative humidity (RH) (85 ± 5 and 45 ± 5 %). Egg development was microscopically examined at days +7, +15, and +30. In addition, in order to assess the vitality of eggs maintained at 5 ± 1 °C for 30 days, these latter cultures were moved, placed at 20 ± 1 °C and 85 ± 5 % RH, and observed for further 30 and 40 days. The results showed that at 20 ± 1 °C and 85 ± 5 % RH, the totality of eggs completed development in 30 days, while about 26 and 70 % of eggs were already fully developed after 7 and 15 days, respectively. No egg development occurred after 30 days at 5 ± 1 °C, while 100 % of eggs placed at 5 ± 1 °C for 30 days and then moved at 20 ± 1 °C and 85 ± 5 % RH for further 40 days were found fully developed.

International audience; Brucella spp. was isolated from brain, lung and intestinal lymph nodes of a dead adult male striped dolphin (Stenella coeruleoalba) found stranded on the Tyrrhenian coast (Tuscany, Italy) of the Mediterranean Sea in February 2012. Brucella spp. was associated with moderate to severe lesions of meningoencephalitis. A co-infection by Toxoplasma gondii was also demonstrated at brain level by means of molecular and histopathologic methods. The Brucella isolate was further characterized based on a fragment-specific polymerase chain reaction (PCR) approach, consisting of a set of five specific PCRs, targeting specific chromosomal IS711 locations for marine mammal Brucellae, as described previously. The isolate was thus classified as Brucella ceti I; V fragment-positive (or B. ceti dolphin type), according to previous studies. Multi Locus Sequence Analysis demonstrated that the isolate belongs to Sequence Type 26, while omp2 (omp2a and omp2b genes) sequence analysis further confirmed the isolate belonged to this group of strains. This is the first report of Brucella spp. from marine mammals in the Mediterranean Sea, and represents a further observation that this strain group is associated with hosts of the Family Delphinidae, and particularly with the striped dolphins, also in the Mediterranean area, thus constituting a further biological hazard of concern for this vulnerable subpopulation.

The aim of the present study was to investigate the in vitro and in vivo acaricidal effects of an essential oil of Cinnamomum zeylanicun leaves on Psoroptes cuniculi, a mange mite. In vitro, 2.5 ml of the essential oil diluted at different concentrations, from 10% to 0.03%, in paraffin oil were added to Petri dishes containing all motile stages of P. cuniculi. Mites mortality observed in these dishes was compared with that observed in untreated and treated (AcaCerulen R) control plates. In vivo, one group of six P. cuniculi infected rabbits was topically treated two times at seven days interval with two ml of the essential oil at the concentration of 2.5% in paraffin oil and compared with untreated and treated (AcaCerulen R) control groups of six rabbits each. After 24 h of contact, all concentrations of essential oil between 0.10 and 10% showed a good in vitro acaricidal efficacy if compared with the untreated controls (p

The inhibitory activity of mangiferin (50 mg/kg/die and 100 mg/kg/die) on Cryptosporidium parvum was evaluated in a neonatal mouse model and its activity was compared with that of paromomycin (100 mg/kg/die). At 4 days of age, neonatal Swiss conventional outbred mice were experimentally infected by oral administration of 10(4) oocysts/animal of C. parvum and treated orally for 10 consecutive days, starting 7 days after the experimental infection. One group of mice was left untreated. To evaluate the efficacy of mangiferin, from euthanised mice, 3-mum-thick tissue sections of the intestine were stained with haematoxylin-eosin and periodic acid Schiff. Immunohistochemistry was also used by employing a monoclonal anti-C. parvum antibody. Oocysts were counted and results were expressed as mean oocysts number/intestine. Results obtained show that mangiferin at 100 mg/kg/die has a significant anticryptosporidial activity and that its activity is similar to that showed by the same dose (100 mg/kg/die) of paromomycin. However, both mangiferin and paromomycin were not able to completely inhibit intestinal colonization of C. parvum but only to reduce it. This reduction was calculated at over 80% for both mangiferin and paromomycin with respect to the untreated control. A significant activity was found also for mangiferin at 50 mg/kg/die only after the end of treatment.

The bacterium Serratia marcescens isolated from surface-sterilised Psoroptes cuniculi was found sensitive to the antibiotic Amikacin. Mites placed in this antibiotic for 48-72 h and then washed by centrifugation were found to be alive and S. marcescens-free. Two experimental infestations were undertaken in order to verify the ability of the S. marcescens-free mites to infect and to give ear skin lesions in healthy rabbits and to evaluate the differential ability of the S. marcescens-free and S. marcescens-infected mites to give ear skin lesions. All rabbits were found to be infested, but only rabbits infested with S. marcescens-free mites presented crusts in their ears, whereas mites and/or eggs were only detected in the ear cerumen of all rabbits infested with S. marcescens-infected mites. S. marcescens was isolated only from P. cuniculi mites taken from these latter rabbits. Results indicate that P. cuniculi mites do not need S. marcescens to live and to be able to infest a healthy rabbit. In addition, S. marcescens was not isolated from eggs and newly born larvae of S. marcescens-infected P. cuniculi, demonstrating that in a population of P. cuniculi this bacterium is not transmitted transovarially.