Description of Babesia galileei sp. nov. A Piroplasmid species causing severe disease in domestic cats.

BACKGROUND: Babesiosis is a tick-borne infection caused by piroplasmid protozoa and associated with anemia and severe disease in humans, domestic animals and wildlife. Domestic cats are infected by at least six Babesia spp. that cause clinical disease. METHODS: Infection with a piroplasmid species was detected by microscopy of stained blood smears in three sick cats from Israel. Genetic characterization of the piroplasmid was performed by PCR amplification of the 18S rRNA, cytochorme B (CytB) and heat shock protein 70 (HSP70) genes and the internal transcribed spacer (ITS) locus, DNA sequencing and phylogenetic analysis. In addition, Haemaphysalis adleri ticks collected from two cats were analyzed by PCR for piroplasmids. RESULTS: The infected cats presented with anemia and thrombocytopenia (3/3), fever (2/3) and icterus (1/3). Comparison of gene and loci sequences found 99-100% identity between sequences amplified from different cats and ticks. Constructed phylogenetic trees and DNA sequence comparisons demonstrated a previously undescribed Babesia sp. belonging to the Babesia sensu stricto (clade X). The piroplasm forms detected included pear-shaped merozoite and round-to-oval trophozoite stages with average sizes larger than those of Babesia felis, B. leo and B. lengau and smaller than canine Babesia s.s. spp. Four of 11 H. adleri adult ticks analyzed from cat # 3 were PCR positive for Babesia sp. with a DNA sequence identical to that found in the cats. Of these, two ticks were PCR positive in their salivary glands, suggesting that the parasite reached these glands and could possibly be transmitted by H. adleri. CONCLUSIONS: This study describes genetic and morphological findings of a new Babesia sp. which we propose to name Babesia galileei sp. nov. after the Galilee region in northern Israel where two of the infected cats originated from. The salivary gland PCR suggests that this Babesia sp. may be transmitted by H. adleri. However, incriminating this tick sp. as the vector of B. galilee sp. nov. would require further studies.

Prevalence and risk factors of Toxoplasma gondii infection in dairy cattle from the Western Region of Thailand.

In total, 901 dairy cow sera and data were collected from 51 farms in Nakhon Pathom, Ratchaburi and Kanchanaburi provinces (Western Region of Thailand). Serum samples were processed via the multispecies ELISA method to detect IgG antibodies against Toxoplasma gondii infection. The results demonstrated that the calculated true prevalence was 1.48% (95% CI, 0.64-2.75%) for the individual-level and 29.41% (95% CI, 18.71-43%) for the farm-level. The univariate risk factor analysis showed that the number of total owned cats, the presence of stray cats, and the frequency of cleaning per day were significant factors (p < 0.2). These three factors were subjected to logistic regression analysis, and the results revealed that the frequency of cleaning farms per day was a potential risk factor for T. gondii-seropositive farms (OR = 2.745, 95% CI, 1.15-8.69, p = 0.02). The frequency of cleaning might increase the T. gondii oocyst distribution within the barn area, thus increasing the possibility of infection. Our findings show that T. gondii continues to circulate in the dairy cow population in the western part of Thailand. The presence of cats on farms was not found to be associated with T. gondii infection, but the high frequency of cleaning the floor was, and contributed to the potential risk of infection.

Title: Prévalence et facteurs de risque de l'infection à Toxoplasma gondii chez les bovins laitiers de la région occidentale de la Thaïlande. Abstract: Au total, 901 sérums de vaches laitières et des données ont été collectés dans 51 fermes des provinces de Nakhon Pathom, Ratchaburi et Kanchanaburi (région occidentale de la Thaïlande). Les échantillons de sérum ont été traités via la méthode ELISA multi-espèces pour détecter les anticorps IgG contre l'infection à Toxoplasma gondii. Les résultats ont démontré que la prévalence réelle calculée était de 1,48 % (IC à 95 %, 0,64­2,75 %) au niveau individuel et de 29,41 % (IC à 95 %, 18,71­43 %) au niveau des exploitations. L'analyse factorielle a montré que le nombre total de chats possédés, la présence de chats errants et la fréquence quotidienne de nettoyage étaient des facteurs significatifs (p < 0,2). Ces trois facteurs ont été soumis à une analyse de régression logistique et les résultats ont révélé que la fréquence quotidienne de nettoyage des exploitations était un facteur de risque potentiel pour les exploitations séropositives à T. gondii (OR = 2,745, IC à 95 % = 1,15­8,69, p = 0,02). La fréquence du nettoyage pourrait favoriser la répartition des oocystes de T. gondii dans les étables, augmentant ainsi le risque d'infection. Nos résultats indiquent que T. gondii continue de circuler dans la population de vaches laitières de l'ouest de la Thaïlande. La présence de chats dans les fermes n'a pas été associée à l'infection à T. gondii, mais la fréquence élevée du nettoyage du sol l'était et contribuait au risque potentiel d'infection.

Feline immunodeficiency virus, feline leukemia virus and Leishmania spp. prevalence in cats from shelters in Mato Grosso do Sul, Brazil.

Diseases such as those caused by feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) represent health problems for cats. Feline leishmaniasis (FL) has been reported in several cities across the country. The objective was to carry out a clinical-epidemiological and laboratory study of FIV, FeLV and FL in cats from shelters in Dourados, Mato Grosso do Sul, Brazil. Blood samples and swabs from the conjunctival and nasal mucosa were obtained from 75 cats, from four animal shelters. Serology for FIV and FeLV was performed. For Leishmania, polymerase chain reaction (PCR) was performed on blood, conjunctiva and nasal mucosa. In the immunochromatographic serological test, seven cats tested positive for FIV and none for FeLV. No samples was positive in PCR for Leishmania. The study showed that despite the presence of human and canine leishmaniasis in the studied region, Leishmania spp. were absent in the cats studied. To avoid an increase in contagion in shelters, it is essential isolate cats with FIV.

Island hitchhikers: pathogen agents of Madeira and Azores ticks.

Ticks are blood-sucking arthropods that can transmit pathogens to their host. As insular ecosystems can enhance tick-host interactions, this study aimed to understand tick diversity, pathogen presence, and their respective associations in the Azores and Madeira archipelagos. Unfed or partially engorged ticks (n = 120) were collected from 58 cats and dogs in the Azores (n = 41 specimens) and Madeira (n = 79 specimens) from November 2018 to March 2019. Vector identification was based on morphology and molecular criteria. For pathogen sequencing, 18S gene fragment for Babesia/Hepatozoon and gltA for Rickettsia were performed. Sequence data was explored using BLAST and BLAST and phylogenetic inference tools. In the Azores, Ixodes hexagonus, I. ventalloi, and Rhipicephalus sanguineus (n = 6; 14.6%, n = 6; 14.6%, and n = 29; 70.7% respectively) were found and in Madeira I. ricinus and R. sanguineus (n = 78, 98.7%; and n = 1, 1.3%; respectively) were identified. Tick COI markers confirmed species highlighting confirmation of R. sanguineus s.s. and genotype A of I. ventalloi. In the Azores Islands, the detected Rickettsia massiliae was linked to R. sanguineus (dogs and cats) and I. hexagonus (dogs), and in Madeira Island, R. monacensis (dogs) and Hepatozoon silvestris (cats) were found associated with I. ricinus. Further, I. ventalloi presence in the Azores expands west its known range, and Hepatozoon silvestris in Madeira may suggest that I. ricinus could have a role as a potential vector. Finally, as R. massiliae and R. monacensis presence underlines public health risks, surveillance by health authorities is crucial as pathogen-tick interactions may drive disease spread, therefore monitoring remains pivotal for disease prevention.

Molecular-phylogenetic investigation of trichomonads in dogs and cats reveals a novel Tritrichomonas species.

BACKGROUND: Trichomonosis is a common infection in small animals, mostly manifesting in gastrointestinal symptoms such as diarrhea. Although oral trichomonads are also known, the species found colonizing the large intestine are more frequently detected protozoa. METHODS: In the present study, four wildcats, 94 domestic cats, and 25 dogs, originating from 18 different locations in Hungary, were investigated for the presence of oral and large intestinal trichomonads based on the 18S rRNA gene and ITS2. RESULTS: All oral swabs were negative by polymerase chain reaction (PCR). However, Tritrichomonas foetus was detected in a high proportion among tested domestic cats (13.8%) and dogs (16%), and Pentatrichomonas hominis only in two domestic cats. In addition, a novel Tritrichomonas genotype was identified in one cat, probably representing a new species that was shown to be phylogenetically most closely related to Tritrichomonas casperi described recently from mice. All positive dogs and half of the positive cats showed symptoms, and among cats, the most frequent breed was the Ragdoll. CONCLUSIONS: With molecular methods, this study evaluated the prevalence of oral and intestinal trichomonads in clinical samples of dogs and cats from Hungary, providing the first evidence of T. foetus in dogs of this region. In contrast to literature data, P. hominis was more prevalent in cats than in dogs. Finally, a hitherto unknown large intestinal Tritrichomonas species (closely related to T. casperi) was shown to be present in a cat, raising two possibilities. First, this novel genotype might have been a rodent-associated pseudoparasite in the relevant cat. Otherwise, the cat was actually infected, thus suggesting the role of a predator-prey link in the evolution of this trichomonad.

Dipylidiasis cases in Japan-an update by literature survey.

Dipylidium caninum is a cosmopolitan parasite of companion animals such as dogs and cats. Accidental infection in humans occur mostly in children. Although considerable number of cases were reported from Europe and the Americas, case reports of this zoonotic disease are rather scarce from Asian countries. The aim of this study is to report the results of literature survey on dipylidiasis cases in humans in Japan. Conclusively, we have found a total of 17 cases since the first case report in from Aichi Prefecture in 1925.

Gastrointestinal parasites of owned cats in three districts of Central Ethiopia: Prevalence and risk factors.

This present, cross-sectional study investigated gastrointestinal parasites in owned cats was carried out across Bishoftu, Dukem, and Addis Ababa city and towns in Central Ethiopia from February 2022 to April 2023. The primary objectives were to estimate the prevalence and assess the risk factors associated with these parasites. A total of 213 faecal samples were collected and processed using flotation and McMaster techniques. An overall prevalence of 34.7% (74/213) of gastrointestinal parasites was recorded in the examined cats. The cats harboured nematode (14.6%), cestode (12.7%), and protozoan (5.6%) parasites in single (32.9%) and mixed (1.9%) species. Specifically, Toxocara cati, Dipylidium caninum, Taenia spp., Giardia spp., and Physaloptera spp. were the more frequently identified parasites, with prevalences of 9.4%, 8.9%, 3.8%, 2.8%, and 2.8%, respectively. Statistically significant differences (P < 0.05) were observed in the prevalence of gastrointestinal parasites among cats of different sexes, ages, and feeding conditions. Thus, higher prevalence rates were observed in females (49.1%, OR = 2.3), young cats (70%, OR = 4.7), and those consuming uncooked food (40.5%, OR = 3.6). Faecal consistency also showed a statistically significant association (P < 0.05) with the presence of gastrointestinal parasites. However, no significant differences were noted in the prevalence among cats of different housing conditions, seasons, or agro-ecological zones. Cats sharing shelter with families had the highest prevalence of gastrointestinal parasites (52.2%). In conclusion, this study uncovered a notably high occurrence of gastrointestinal parasites in cats in the surveyed regions, emphasizing the imperative of adopting a One Health strategy to address the zoonotic parasites found in cats, which pose significant public health risks.

Evaluation of Parasight All-in-One system for the automated enumeration of helminth ova in canine and feline feces.

BACKGROUND:  Digital imaging combined with deep-learning-based computational image analysis is a growing area in medical diagnostics, including parasitology, where a number of automated analytical devices have been developed and are available for use in clinical practice. METHODS: The performance of Parasight All-in-One (AIO), a second-generation device, was evaluated by comparing it to a well-accepted research method (mini-FLOTAC) and to another commercially available test (Imagyst). Fifty-nine canine and feline infected fecal specimens were quantitatively analyzed by all three methods. Since some samples were positive for more than one parasite, the dataset consisted of 48 specimens positive for Ancylostoma spp., 13 for Toxocara spp. and 23 for Trichuris spp. RESULTS: The magnitude of Parasight AIO counts correlated well with those of mini-FLOTAC but not with those of Imagyst. Parasight AIO counted approximately 3.5-fold more ova of Ancylostoma spp. and Trichuris spp. and 4.6-fold more ova of Toxocara spp. than the mini-FLOTAC, and counted 27.9-, 17.1- and 10.2-fold more of these same ova than Imagyst, respectively. These differences translated into differences between the test sensitivities at low egg count levels (< 50 eggs/g), with Parasight AIO > mini-FLOTAC > Imagyst. At higher egg counts Parasight AIO and mini-FLOTAC performed with comparable precision (which was significantly higher that than Imagyst), whereas at lower counts (> 30 eggs/g) Parasight was more precise than both mini-FLOTAC and Imagyst, while the latter two methods did not significantly differ from each other. CONCLUSIONS: In general, Parasight AIO analyses were both more precise and sensitive than mini-FLOTAC and Imagyst and quantitatively correlated well with mini-FLOTAC. While Parasight AIO produced lower raw counts in eggs-per-gram than mini-FLOTAC, these could be corrected using the data generated from these correlations.

A seven-legged tick - anomalous Ixodes ricinus female (Acari: Ixodidae) from Poland.

Morphological anomalies are considered a rare phenomenon among natural tick populations. New cases of abnormalities in ticks are being described, such as body assymetries, nanism, gynandromorphism and limb malformations. The tick removed from a cat was morphologically identified to species and developmental stage. The time of feeding on the host was determined. The specimen was tested using PCR and Real-Time PCR methods for the presence of the common tick-borne pathogens: Anaplasma phagocytophilum, Babesia spp, Borrelia spp., Neoehrlichia mikurensis, Rickettsia spp. For visualisation of the anomalous structures, scanning electron microscopy (SEM) was performed. The tick was identified as a slightly engorged adult female of I. ricinus exhibiting ectromely of leg I on the left side of the idiosoma. The specimen was tested positive for two medically important pathogens: A. phagocytophilum and N. mikurensis. The case report describes a rare case of a morphological anomaly in an I. ricinus tick from Poland.

Detection of rat lungworm (Angiostrongylus cantonensis) infection by real-time PCR from the peripheral blood of animals: a preliminary study.

Rat lungworm disease or neuroangiostrongyliasis is a cerebral parasitic infection that affects humans and animals alike. Its clinical signs and symptoms can range from mild self-resolving to serious life-threatening conditions. Studies suggest therapeutic interventions during the early stages of infection to be more effective than in later stages. However, early diagnosis of infection is usually problematic without the knowledge of exposure and/or detection of the parasite's DNA or antibody against the parasite in the cerebrospinal fluid. This requires a lumbar puncture, which is an invasive procedure that generally requires hospitalization. This study evaluates an affordable and less invasive alternative to detect parasitic DNA by PCR from the peripheral blood of potentially infected animals. Blood samples from 58 animals (55 dogs and 3 cats) with clinical suspicion of infection were submitted to our lab between February 2019 and August 2022 by local, licensed veterinarians. DNA was extracted from whole blood, plasma, serum, and/or packed cells using the Qiagen DNeasy Blood & Tissue Kit as per the manufacturer's protocol. All 58 animals were tested by real-time PCR using the AcanITS1 assay and 32 of these animals (31dogs; 1 cat) were also tested using the AcanR3990 assay. The PCR results for both assays were classified into strongly positive > positive > weakly positive > negative, and equivocal for ambiguous results, based on the strength of the signal. The percent infection detected using the AcanITS1 and AcanR3990 assays was 12.72% (7/55) and 20.68% (6/29), respectively. The overall percent infection detected was 34.37% (11/32), with only two animals testing positive by both assays. The three cats involved in this study tested negative by both assays. These results are promising and warrant further investigations to increase sensitivity including variables that might affect detection in the blood, such as parasite load, and laboratory methodologies.

Feline sarcoptic mange in Poland: A case series of three cats.

BACKGROUND: Sarcoptic mange is rare in cats. The main symptoms reported in cases of feline sarcoptic mange include crusty lesions and pruritus, although these may vary in severity among individuals. OBJECTIVES: This report describes three cats infested with Sarcoptes scabiei, all presenting with pruritus and excoriation. METHODS: The diagnosis was confirmed by microscopic observation of skin scrape samples. RESULTS: All three cats were treated successfully using moxidectin and imidacloprid, selamectin and ivermectin, respectively. CONCLUSIONS: The clinical presentation of feline scabies appears to be more variable in cats than in dogs. Infestation with S. scabiei should be considered a differential diagnosis for cats presenting with pruritic inflammatory skin disease.

Molecular, morphological and histopathological evidence of Spirometra mansoni in wild and domestic animals from Costa Rica.

Spirometra mansoni is a diphyllobothroid cestode and one of the causing agents of sparganosis, a zoonotic foodborne and waterborne infection in humans. This parasite has an indirect life cycle with domestic and wild canids or felids as definitive hosts. The last report of S. mansoni in Costa Rica was done in 2004 by morphological assessment of worms, whereas molecular evidence of this species was obtained recently in the Americas. Herein, we present seven cases of spirometrosis in four dogs, three cats and a coyote from different regions of Costa Rica occurring in a time span of a year. Dog cases presented vomiting, hyporexia, lethargy and diarrhea, whereas cats were mostly asymptomatic. Moreover, the coyote was found with Spirometra sp. proglottids incidentally. Cytochrome oxidase subunit 1 (cox1) sequences of eggs or proglottids derived from all cases were analyzed with a Bayesian Inference phylogenetic tree and a haplotype network. These analyses showed the clustering of S. mansoni from Costa Rica with other sequences derived from Asia and America. Moreover, cox1 sequences clustered in two separate haplotypes, suggesting the high genetic diversity of the species. The present cases represent the first molecular evidence of the parasite in Central America; thus, extending its known range in the American continent.

Geoepidemiology, seroprevalence and factors associated with Toxoplasma gondii infection in domicilied cats from Paraíba (Brazil).

Toxoplasma gondii is a parasite responsible for toxoplasmosis, an emerging and often neglected zoonosis in South America, particularly Brazil. Felines, the only definitive hosts, excrete oocysts in their feces, potentially infecting all homeotherms. Domestic cats are primarily responsible for contaminating human environments with these oocysts. Monitoring their populations is therefore essential to ensure proper toxoplasmosis prophylaxis. The aim of this study was to estimate the prevalence of T. gondii and exposure factors in a population of owner cats in the city of João Pessoa, Paraíba, Brazil. A total of 267 blood samples were collected from domestic cats aged between 1 and 15 years and tested with an immunofluorescence antibody test. The seroprevalence of antibodies against T. gondii was only 17.22% (46/267 individuals). This result therefore suggests a low contribution of domestic cats to T. gondii contamination of the urban environment. The cats' age and living environment were identified as risk factors for cat exposure to T. gondii.

Title: Géoépidémiologie, séroprévalence et facteurs associés à l'infection à Toxoplasma gondii chez les chats domiciliés à Paraíba (Brésil). Abstract: Toxoplasma gondii est le parasite responsable de la toxoplasmose, une zoonose émergente et souvent négligée en Amérique du Sud, notamment au Brésil. Les félins, seuls hôtes définitifs, excrètent des oocystes dans leurs selles, infectant potentiellement tous les homéothermes. Les chats domestiques sont les premiers responsables de la contamination des environnements humains avec ces oocystes. La surveillance de leurs populations est donc essentielle pour garantir une prophylaxie adéquate contre la toxoplasmose. Le but de cette étude était d'estimer la prévalence de T. gondii et les facteurs d'exposition dans une population de chats domestiques de la ville de João Pessoa, Paraíba, Brésil. Au total, 267 échantillons de sang ont été prélevés sur des chats domestiques âgés de 1 à 15 ans et testés avec un test d'immunofluorescence des anticorps. La séroprévalence des anticorps contre T. gondii n'était que de 17,22 % (46/267 individus). Ce résultat suggère donc une faible contribution des chats domestiques à la contamination du milieu urbain par T. gondii. L'âge et le milieu de vie des chats ont été identifiés comme facteurs de risque d'exposition du chat à T. gondii.

Hepatozoon spp. in stray cats from the metropolitan area of Rio de Janeiro, Brazil.

In the last few years, the number of studies on feline hepatozoonosis has increased, but our knowledge on the actual species of Hepatozoon and/or different genotypes affecting felines is still incipient. At least three species, namely Hepatozoon felis, H. canis, and H. silvestris, have been isolated from domestic cats in various countries. Additionally, there are indications that other species and genotypes may affect felines in given geographic areas. This study was carried out to investigate the occurrence of Hepatozoon spp. in cats from Niterói, a municipality within the metropolitan area of Rio de Janeiro, Brazil. Individual blood samples were collected from 28 cats enrolled in a spaying/castration program. DNA was extracted from all samples and subjected to sequencing specific for Hepatozoon spp. DNA of H. felis was found in 21/28 cats (75%), and four genetic polymorphisms never described thus far were detected. This is the first report of H. felis in cats living in the State of Rio de Janeiro, and the present data confirm that H. felis is a species complex encompassing different genotypes circulating within cat populations. Further studies are warranted to investigate whether different genotypes have different biology or pathogenicity for felids.

Title: Hepatozoon spp. chez les chats errants de la zone métropolitaine de Rio de Janeiro, Brésil. Abstract: Au cours des dernières années, le nombre d'études sur l'hépatozoonose féline a augmenté, mais nos connaissances sur les espèces d'Hepatozoon et/ou différents génotypes affectant les félins sont encore naissantes. Au moins trois espèces, à savoir Hepatozoon felis, H. canis et H. silvestris, ont été isolées chez des chats domestiques dans divers pays. De plus, il semble que d'autres espèces et génotypes puissent affecter les félins dans des zones géographiques données. Cette étude a été réalisée pour étudier la présence d'Hepatozoon spp. chez des chats de Niterói, une municipalité de la zone métropolitaine de Rio de Janeiro, au Brésil. Des échantillons de sang ont été prélevés individuellement sur 28 chats d'un programme de castration. L'ADN a été extrait de tous les échantillons et soumis à un séquençage spécifique de Hepatozoon spp. L'ADN de H. felis a été trouvé chez 21 chats sur 28 (75%) et quatre polymorphismes génétiques, jamais décrits jusqu'à présent, ont été détectés. Il s'agit du premier signalement de H. felis chez des chats vivant dans l'État de Rio de Janeiro et les données actuelles confirment que H. felis est un complexe d'espèces englobant différents génotypes circulant au sein des populations de chats. Des études supplémentaires sont nécessaires pour déterminer si les différents génotypes ont une biologie ou une pathogénicité différente pour les félidés.

A case of mistaken identity: a systematic review, meta-analysis, and reinvestigation of hemotropic Mycoplasma spp. infection in Ctenocephalides felis (cat flea).

BACKGROUND: Feline-associated hemotropic Mycoplasma (hemoplasmas) are believed to be transmitted by two primary mechanisms: (1) direct transmission via fighting and (2) vector-borne transmission by the cat flea (Ctenocephalides felis). While the efficiency of transmission by C. felis appears low, most manuscripts focus on the prevalence of hemoplasmas in wild-caught fleas and report either a very low (< 3%) or a high (> 26%) prevalence. Therefore, we aimed to assess the influence of sample processing and PCR methods on C. felis hemoplasma infection prevalence. METHODS: A systemic review of PubMed articles identified 13 manuscripts (1,531 fleas/flea pools) that met the inclusion criteria (performed PCR for >1 hemoplasma on C. felis collected from cats). Risk of bias was assessed utilizing the ROBINS-E tool. Meta-analysis performed in R of these manuscripts found that not washing samples and a common set of 16S rRNA primers first published in Jensen et al. 2001 were associated with increased hemoplasma prevalence. To evaluate the influence of washing on newly collected fleas, we assessed the hemoplasma status of 20 pools of 5 C. felis each, half of which were washed and half not washed. RESULTS: Flea washing did not influence the detection of hemoplasma but instead amplified Spiroplasma. To assess non-specific amplification with the Jensen et al. 2001 primers, 67 C. felis samples (34% previously reported hemoplasma infected) were subject to PCR and sequencing. By this method, hemoplasma was detected in only 3% of samples. In the remaining "hemoplasma infected" fleas, PCR amplified Spiroplasma or other bacteria. CONCLUSIONS: Therefore, we concluded that hemoplasma infection in C. felis is rare, and future flea prevalence studies should sequence all positive amplicons to validate PCR specificity. Further investigation of alternative methods of feline-associated hemoplasma transmission and the ability of C. felis to maintain hemoplasma infection is necessary.

Awareness of parasitic zoonotic diseases among pet owners in Cairo, Egypt.

Egyptians are becoming more interested in owning and raising pets; however, most of them lack essential awareness about the risk of zoonotic parasites that could be transmitted. The objective of the present investigation was to evaluate the degree of awareness Egyptian pet owners possess concerning zoonotic parasitic diseases, the risk of transmission, and preventative measures. A cross-sectional study was conducted using an e-survey. Among 246 pet owners, 64.2% (158) were females, and 67.9% (167) belonged to the 20-30 age group. The majority, 78.9% (194), were raising cats. Only 13.8% (34) visited the veterinarian regularly, with significantly higher results among dog owners (p < 0.05). Only 31.3% (77) participants were regularly deworming their pets, and 19.9% (49) were giving their pets prophylaxis against ectoparasites, with significantly higher results among those who visited the veterinarian regularly (p < 0.0001) and among dog owners (p < 0.05). Only 54.1% (133) had heard about the term "zoonoses" before, and about 8.9% (22) of participants showed a history of zoonotic parasitic diseases, with significantly higher results among those who allowed their animals to play with other animals of neighbors and friends (p < 0.05). The obtained results concluded that the surveyed group had a relatively good degree of knowledge regarding pets as a source of zoonotic illness; raising pet owners' awareness regarding the importance of routine medical examinations and minimizing the contact of pets with other stray animals is essential.

Cytauxzoonosis in Indiana, USA: a case series of cats infected with Cytauxzoon felis (2018-2022).

CASE SERIES SUMMARY: This case series describes six cases involving seven cats naturally infected with Cytauxzoon felis in Indiana, USA. Medical records were retrospectively reviewed and all available information on signalment, history, clinical and diagnostic findings, treatment, outcome and pathology was reported. Cats infected with C felis were domestic shorthairs, were aged between 2 and 9 years and all but one of the cats were male. The seven infected cats originated from five counties in southwestern Indiana. Six of seven cats were found to have acute cytauxzoonosis based on clinical signs, gross pathologic lesions, observation of C felis in tissues and/or detection of C felis DNA. One cat was identified as a subclinical survivor cat with no known clinical history of cytauxzoonosis. RELEVANCE AND NOVEL INFORMATION: The reported cases are the first confirmed reports of acute and chronic cytauxzoonosis in cats from Indiana and document an expansion in the range of C felis. Veterinary practitioners in Indiana should consider infection with C felis as a differential diagnosis for cats that present with fever, inappetence, lethargy, depression, dehydration, dyspnea, hemolytic crisis, anorexia or icterus. Administration of approved acaricides to cats currently offers the best protection and control against C felis infection.

Microscopic and molecular prevalence and associated risk factors with Toxocara and Blastocystis infection in dogs and cats in Mitidja, Algeria.

Domestic dogs and cats can serve as a source of environmental contamination with Toxocara spp. and Blastocystis spp., and this represents a neglected public and veterinary health problem. We assessed the microscopic and molecular prevalence of these species in a locality in Algeria and identified the associated risk factors. The faeces of 225 dogs and 78 cats were collected in Mitidja between March and July 2022. The samples were analysed by coproscopy and by polymerase chain reaction (PCR) targeting the Internal Transcribed Spacer 2 (ITS2) and Small Subunit Ribosomal (SSU-RNA) of T. canis and Blastocystis spp. respectively. The overall microscopic prevalence of Toxocara spp. in dogs and cats was 9.78 ± 1.98% and 12.82 ± 7.42%, respectively. The rate of Blastocystis spp. was 15.11 ± 2.39% and 15.38 ± 4.08% in dogs and cats, respectively while the molecular prevalence of T. canis in dogs was 4.89 ± 1.44% and in cats 1.28 ± 1.27%; the prevalence of Blastocystis spp. was 41.78 ± 3.29% and 34.62 ± 5.39% in dogs and cats, respectively. Phylogenetic and phylogeographic analyses identified the presence of the H1 subtype of T. canis in dogs, and the ST1 subtype of Blastocystis in dogs and cats. Dogs with clinical signs were more likely to be infected with T. canis (OR 6.039, P < 0.05) than healthy dogs. This study demonstrates that dogs and cats are carriers of Toxocara spp. and Blastocystis spp. and are therefore a source of environmental contamination. Veterinarians and human health professionals should work together to implement control strategies as part of a "One Health" approach to improving animal health and reducing the risk of transmission to humans.

RPA-CRISPR/Cas12a-LFA combined with a digital visualization instrument to detect Toxoplasma gondii in stray dogs and cats in Zhejiang province, China.

Toxoplasma gondii, which causes toxoplasmosis, is prevalent in warm-blooded animals, such as cats, dogs, and humans. T. gondii causes economic losses to livestock production and represents a potential risk to public health. Dogs and cats are common hosts in the epidemiology of toxoplasmosis. The current molecular diagnostic tools for T. gondii infection require high technical skills, a laboratory environment, and complex instruments. Herein, we developed a recombinase polymerase amplification (RPA)-clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 12a (Cas12a) assay to detect T. gondii. The lowest limit of detection of the assay was 31 copies/µL for the T. gondii B1 gene. In addition, we established a visual RPA-CRISPR/Cas12a lateral flow band assay (RPA-CRISPR/Cas12a-LFA) combined with a digital visualization instrument, which minimized the problem of false-negative results for weakly positive samples and avoided misinterpretation of the results by the naked eye, making the LFA assay results more accurate. The assay established in this study could identify T. gondii within 55 min with high accuracy and sensitivity, without cross-reaction with other tested parasites. The developed assay was validated by establishing a mouse model of toxoplasmosis. Finally, the developed assay was used to investigate the prevalence of T. gondii in stray cats and dogs in Zhejiang province, Eastern China. The positive rates of T. gondii infection in stray cats and dogs were 8.0% and 4.0%, respectively. In conclusion, the RPA-CRISPR/Cas12a-LFA is rapid, sensitive, and accurate for the early diagnosis of T. gondii, showing promise for on-site surveillance. IMPORTANCE: Toxoplasma gondii is a virulent pathogen that puts millions of infected people at risk of chronic disease reactivation. Hosts of T. gondii are distributed worldwide, and cats and dogs are common hosts of T. gondii. Therefore, rapid diagnosis of early T. gondii infection and investigation of its prevalence in stray dogs and cats are essential. Here, we established a visual recombinase polymerase amplification-clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 12a-assay combined with a lateral flow band assay and a digital visualization instrument. Detailed analyses found that the assay could be used for the early diagnosis of T. gondii without false-negative results. Moreover, we detected the prevalence of T. gondii in stray cats and dogs in Zhejiang province, China. Our developed assay provides technical support for the early diagnosis of T. gondii and could be applied in prevalence surveys of T. gondii in stray dogs and cats.

Efficacy of Bravecto® Plus spot-on solution for cats (280 mg/ml fluralaner and 14 mg/ml moxidectin) in the prevention of feline Aelurostrongylus abstrusus infection evaluated in a multi-diagnostic approach.

BACKGROUND: Aelurostrongylus abstrusus is one of the most important respiratory nematodes of felines. Infections may lead to respiratory clinical signs with varying severity or even death, emphasizing the need for preventive treatment of cats with outdoor access to circumvent patent infections. METHODS: Therefore, the preventive efficacy of a spot-on formulation of 280 mg/ml fluralaner and 14 mg/ml moxidectin (Bravecto® Plus spot-on solution for cats, MSD) against A. abstrusus was evaluated in a negative controlled, randomized and partially blinded efficacy study with 28 purpose-bred cats in a non-terminal design. In three different treatment regimes, the minimum recommended dose of 40 mg fluralaner and 2.0 mg moxidectin/kg bodyweight (BW) was administered once at 12, 8 or 4 weeks (study group G1, G2 and G3, respectively) prior to experimental infection with 300 third-stage A. abstrusus larvae, while G4 served as placebo-treated control. RESULTS: From 30 to 46 days post infection (dpi; SD 114 to 130), faeces were sampled to monitor first-stage larvae (L1) excretion for efficacy determination. Secondary efficacy criteria, including respiratory parameters, serological antibody levels and computed tomography (CT) findings, were assessed once before enrolment (SD -7 to -1) and before infection (SD 75 to 83). After infection, CT evaluation was performed once at 47-50 dpi (SD 131 to 134), and respiratory parameters and antibody levels were regularly assessed twice or once a week, respectively (1 up to 78 dpi, SD 85 up to 162). All animals in the control group excreted L1 by 33-37 dpi and remained positive throughout the study period from 41 to 46 dpi (SD 125 to 130). In the treatment groups, only one animal each of G1 and G2 excreted L1 at two consecutive days, and four cats of G1, two of G2 and three of G3 were positive on single occasions. While the geometric mean (GM) of the maximum number of excreted L1 per 5 g of faeces was 7380.89 in the control group (G4), GMs were significantly lower in the treatment groups with 1.63 in G1, 1.37 in G2 and 0.79 in G3. Thus, based on GMs, the reduction in excreted L1 exceeded 99.9% in all three treatment groups. Based on CT severity scores, all lungs of the animals of the control group showed severe pulmonary changes post infection, whereas lungs of the cats of the treatment groups were either unaltered (4 animals), mildly (11 animals), or moderately altered (5 animals). Moreover, seroconversion was observed in all cats of the control group, but not in those of the treatment groups. CONCLUSIONS: The combination of diagnostic methods used in this non-terminal study yielded coherent and reliable results. A single administration of Bravecto® Plus spot-on solution for cats was well tolerated and effective in the prevention of aelurostrongylosis for at least 12 weeks.