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Felicola subrostratus is the only species of louse affecting domestic cats. Although it is considered a rare ectoparasite of pet cats, it occurs mainly in stray or shelter animals and is sometimes associated with intense pruritus and secondary bacterial infections. The aim of the present study was to evaluate the efficacy of a single dose of the topical formulation of esafoxolaner, eprinomectin and praziquantel (NexGard Combo ® , Boehringer Ingelheim) in cats for the treatment of naturally acquired chewing lice infestation in a multi-site, positive-control, blinded clinical field study. Thirty-one domestic cats presenting natural F. subrostratus infestation were included in the study. The animals had not been treated with any ectoparasiticide within the previous 60 days. After inclusion, each cat was randomly assigned to one of the two groups: group 1, 14 cats treated with NexGard Combo ® or group 2, 17 cats treated with Frontline Combo ® . A clinical evaluation was performed at days 0 (inclusion), 14 and 30 and consisted in scoring the skin lesions and symptoms and scoring the presence of lice. On day 30, all cats from both groups scored 0 for the presence of live lice and no dead lice were found, demonstrating 100% efficacy. The clinical scores significantly improved from day 0 to day 30 in both groups. During the 30 days of surveillance, no reinfestations due to the hatching of eggs were observed and none of the cats had any adverse reactions. Esafoxolaner demonstrated high efficacy for the treatment of F. subrostratus infestation. Felicola subrostratus est la seule espèce de pou affectant les chats domestiques. Bien que ce pou soit rare comme ectoparasite chez les chats de compagnie, il infeste principalement les animaux errants ou en refuge et est parfois associé à un prurit intense et à des infections bactériennes secondaires. Le but de la présente étude était d’évaluer l’efficacité d’une dose unique de la formulation topique d’esafoxolaner, d’éprinomectine et de praziquantel (NexGard Combo ® , Boehringer Ingelheim) chez le chat pour le traitement de l’infestation naturelle par les poux broyeurs dans une étude de terrain multi-sites, randomisée, avec un contrôle positif. Trente et un chats domestiques présentant une infestation naturelle par F. subrostratus ont été inclus dans l’étude. Les animaux n’avaient été traités avec aucun ectoparasiticide au cours des 60 jours précédents. Après inclusion, chaque chat a été assigné au hasard à l’un des deux groupes : groupe 1, 14 chats traités avec NexGard Combo ® , ou groupe 2, 17 chats traités avec Frontline Combo ® . Une évaluation clinique a été réalisée aux jours 0 (inclusion), 14 et 30 et a consisté à un score clinique des lésions et symptômes cutanés et à noter la présence de poux. Au jour 30, tous les chats des deux groupes ont obtenu un score de 0 pour la présence de poux vivants et aucun pou mort n’a été trouvé, démontrant une efficacité de 100 %. Les scores cliniques se sont significativement améliorés du jour 0 au jour 30 dans les deux groupes. Au cours des 30 jours de surveillance, aucune réinfestation due à l’éclosion des œufs n’a été observée et aucun des chats n’a présenté d’effet indésirable. L’esafoxolaner a démontré une efficacité élevée pour le traitement de l’infestation par F. subrostratus .

Tickborne bacterial pathogens have been described worldwide as risk factors for both animal and human health. Spotted fevers caused by Rickettsiae may cause non-specific symptoms, which make clinical diagnosis difficult. The aim of the current study was to evaluate and review the diversity of SFG Rickettsiae in ticks collected in 41 counties in Romania. A total of 2028 questing and engorged ticks collected in Romania belonging to five species were tested by PCR amplification of Rickettsia spp. gltA and 17-D gene fragments: Ixodes ricinus (n = 1128), Dermacentor marginatus (n = 507), D. reticulatus (n = 165), Rhipicephalus rossicus (n = 128) and Haemaphysalis punctata (n = 100). Five Rickettsia species were identified following DNA sequence analysis: R. helvetica, R. monacensis, R. slovaca, R. raoultii, and R. hoogstraalii. The most common species detected was R. monacensis. Moreover, R. hoogstraalii was detected for the first time in Romania and in R. rossicus ticks. The detection of R. raoultii and R. monacensis in questing larvae of Hae. punctata suggests the possible transovarial transmission of these Rickettsia species in ticks. The detection of R. hoogstraalii for the first time in Romania increases the reported SFG Rickettsia diversity in the country.

BackgroundTicks are obligate hematophagous arthropods capable of transmitting a great variety of endemic and emerging pathogens causing diseases in animals and humans.ObjectivesThe aim of this study was to investigate the presence of Bartonella spp., Rickettsia spp., Borrelia burgdorferi sensu lato (s.l.) and Anaplasma phagocytophilum in ticks collected from cattle in Benin and Togo.MethodsOverall, 396 (148 males, 205 females and 43 nymphs) ticks were collected from cattle in 17 districts (Benin and Togo) between 2019 and 2020. Ticks were pooled into groups of 2–6 ticks per pool according to individual host, location, species and developmental stage. The DNA of each pool was extracted for molecular screening.ResultsPCR results revealed that 20 tick pools were positive for Bartonella spp. (Benin and Togo) and 23 tick pools positive for Rickettsia spp. (Benin), while all pools were negative for A. phagocytophilum and B. burgdorferi s.l. Sequence analysis of positive Rickettsia samples revealed the presence of Rickettsia aeschlimannii.ConclusionsThe present study highlights the presence of zoonotic agents in ticks collected from cattle in Benin and Togo. This information will raise awareness of tick-borne diseases among physicians and veterinarians, stimulate further studies to monitor these pathogens, and advise on necessary measures to control the spread of these zoonoses.

Background Trichodectes canis is a small chewing louse found globally that primarily infests dogs. Limited information is available on the efficacy of isoxazolines against infestation with the chewing louse. In the present study, we evaluated the efficacy of afoxolaner, an isoxazoline class compound, in naturally infested domestic dogs. Methods The field study was carried out in Romania. Between September 2021 and December 2021, 43 dogs with confirmed T. canis infestation were included in the study. On the day of the inclusion (day 0), each animal was clinically examined and randomly treated with a control product labeled for use against lice [fipronil-(S)-methoprene combination (Frontline Combo®; Boehringer Ingelheim)] or with the investigational product [chewable tablets containing afoxolaner (NexGard®; isoxazoline)]. Each animal was evaluated for the presence of lice at 15 and 30 days post-inclusion. Results Of the 48 dogs initially included in the study, 43 completed the treatment period [18 in the control group (CG) and 25 in the investigational group (IG)]. At day 14, no living T. canis lice were detected on the dogs in either group. At day 14, dead lice were detected in four dogs in the IG, while eggs were present in two dogs in the IG and in one dog in the CG. At day 30, no lice were detected in either group, while eggs were still present in one dog in the CG. Conclusion These results suggest that afoxolaner is a feasible treatment option against chewing lice in dogs, providing 100% curative efficacy. Graphical Abstract

African swine fever has a significant economic, social and environmental impact due to official regulation of the disease, namely the mass depopulation of all pigs in confirmed outbreaks. The main objective of the present study was to statistically analyze the possible correlation between the number of outbreaks and infected pigs from backyard farms with the altitude, seasonal average annual humidity, and average temperature during spring, summer, autumn and winter, as well as the distance from forests, rivers, and lakes in Romania. The study included all infected backyard pigs with African swine fever virus (n = 7764) and outbreaks (n = 404) that occurred in Romania between 6 February 2020 and 2 March 2021. The number of infected pigs and ASF outbreaks were significantly higher in localities at altitudes between 100 and 500 m, average annual humidity between 60% and 80%, average spring temperature between 10 and 14 °C, average summer temperature between 16 and 22 °C, average autumn temperature between 10 and 13 °C, average winter temperatures between −1 and 2 °C, and at distances of less than 5 km from the forests, less than 15 km from lakes and less than 5 km from the rivers. The number of affected pigs decreased significantly at summer temperatures below 16 °C.

Background The extraordinary Galapagos Islands, with an impressive number of endemic and native species, maintain the interest and curiosity for researchers from all over the world. The native species are known to be vulnerable to new pathogens, cointroduced with their invasive hosts. In the case of invasive parasitic arthropods, their evolutionary success is related to the association with other invasive hosts (such as domestic animals). These associations could become a significant driver of change, as occasionally they can seek another hosts and have the capacity to transmit pathogens between domestic and wild animals. The current study aims to identify the distribution and abundance of canine vector-borne parasites in the Galapagos Islands based on the possibility that some of them could spill over to endemic mammals. Methods A total of 1221 blood samples were randomly collected from privately owned dogs on San Cristóbal, Isabela, Santa Cruz, and Floreana Islands during the years 2021 and 2022. All samples were examined for vector-borne pathogens using the modified Knott’s test and conventional, nested, and multiplex polymerase chain reactions (PCRs), followed by sequencing. Results The PCR and Knott tests confirmed the presence of Dirofilaria immitis (2%, 25/1221) in all islands. While molecular analyses showed heartworm DNA only in dogs from the San Cristóbal (3.3%) and Isabela (2.4%) Islands. Moreover, other pathogens Babesia vogeli (3%, 37/1221) and Hepatozoon canis (0.2%, 2/1221) were detected for the first time by molecular analyses. Conclusions Dogs from Galapagos are hosts to various pathogens, of which some are potentially zoonotic while some other could be spill-over to endemic endangered carnivores, such as sea lions. To understand and limit their impact, long-term surveillance, control, and awareness is needed. Graphical abstract

Background In a countrywide study aiming to update the knowledge on diversity of sand fly species in Romania, a sand fly population was observed in an isolated system of cave microhabitats. The caves are located in the protected area of Canaraua Fetii, Dobrogea region, southeastern Romania. The highest sand fly diversity was recorded in this area between 1968 and 1970. This work presents a study conducted to estimate the seasonal variation of the sand fly species in correlation with the particular environmental factors of the isolated system of cave microhabitats. Methods Sand flies were collected between May and October 2020 from one trapping site of interest in Canaraua Fetii. The trapping site consisted of a cave entrance. CDC miniature light traps and sticky traps were used to collect insects from the exterior walls of the cave entrance. Species identification of collected sand flies was done using morphological keys. Statistical analysis of the trapping and climatic data was performed. Results From all collected sand flies, 99.7% (818/822) were Phlebotomus neglectus , 0.1% (1/822) Ph. balcanicus and 0.2% (2/822) Sergentomyia minuta . Sand fly activity was first observed on 2 July and last on 24 September. A monomodal abundance trend was present, with the peak activity between 16 and 17 July. The analysis of the climatic data showed correlations between the total number of captured sand flies and both average temperature and average relative humidity. The total number of collected specimens was statistically higher when CDC miniature light traps were used compared to sticky traps. The number of females on the sticky traps was significantly higher than the number of males on the same trap type. Compared with the sticky traps, significantly more males were collected by CDC miniature light traps. This is the first record of Se. minuta in Romania after 50 years of no records (despite the trapping effort of the last 5 years in the country). Also, Ph. sergenti , previously present in this location, was not found. Conclusions In the investigated natural habitat, the diversity of the sand fly species appears to have changed, with the predominance of Ph. neglectus instead of Ph. balcanicus and Se. minuta (recorded as the two predominant species in 1968–1970). A monomodal abundance trend was observed as in other regions of the country. The sand fly activity in this particular cave microhabitat appears to be longer than in other regions in Romania. Longer sand fly activity increases the zoonotic risk of various pathogenic species’ transmission, with an impact on public health, as sand flies are important insect vectors. Graphical abstract

Background Apicomplexan haemoparasites are protozoans that infect a variety of domestic and wild animal species, as well as humans. Data regarding haemoprotozoans in domestic cats are limited; therefore, the aim of this study was to assess the occurrence of Babesia spp., Cytauxzoon spp., and Hepatozoon spp. in domestic cats in Romania using molecular tools. Methods Blood samples from 371 domestic cats were screened for the presence of piroplasmids. All samples that yielded a visible band in agarose gels were subsequently tested by specific assays targeting the 18S rDNA of Babesia spp., Cytauxzoon spp., and Hepatozoon spp. Moreover, nested PCR assays targeting mitochondrial genes of Babesia spp. were used for screening of all Babesia spp. 18S rDNA -positive samples. Results From the total number of sampled cats, 19.4% were positive in the PCR assay targeting piroplasmids. Babesia spp. were identified in 15.1% of cats, while 0.5% were positive for Hepatozoon spp. Molecular analyses confirmed the presence of Babesia canis . No samples were positive for Cytauxzoon spp. Conclusions The high infection rates of domestic cats with Babesia spp. and the need for species differentiation highlight the importance of mitochondrial genes as targets for molecular protocols.

This study aimed to evaluate microbial growth, pathogen presence, and histamine production in rainbow trout and mackerel stored on flaked ice over their shelf life. A total of 72 fish samples (rainbow trout and mackerel) were analyzed across four storage intervals (day 1, 3, 9, 12/11) on flaked ice. TVC increased from 2.59 to 5.04 log cfu/g in rainbow trout and from 3.18 to 4.88 log cfu/g in mackerel over the storage period. Significant increases were observed in Pseudomonas, Aeromonas, and Enterobacteriaceae populations, especially after the ninth day. Microbial identification revealed spoilage-associated bacteria, such as Pseudomonas fluorescens and Aeromonas salmonicida, as well as opportunistic pathogens, including Francisella tularensis, Yersinia spp., and Chromobacterium violaceum. Histamine levels rose with storage time but remained below toxic thresholds (<200 mg/kg), peaking at 1.56 mg/kg in trout and 1.87 mg/kg in mackerel. A strong positive correlation was found between TVC and histamine levels (Pearson’s r = 0.85 for trout, 0.82 for mackerel). Proper hygiene and storage are crucial, and consumption is recommended before day 9 of storage on flaked ice. Hygiene measures remain essential to minimize contamination risks and preserve product safety.

Haemoparasites of the genus Babesia infect a wide range of domestic and wild animals. Feline babesiosis is considered endemic in South Africa, while data on Babesia spp. infection in felids in Europe is scarce. Using samples from 51 wild felids, 44 Felis silvestris and 7 Lynx lynx, the study aimed to determine the presence and genetic diversity of Babesia spp. in wild felids in Romania by analyzing the 18S rDNA and two mitochondrial markers, cytochrome b (Cytb) and cytochrome c oxidase subunit I (COI) genes. By 18S rDNA analyses, Babesia spp. DNA was detected in 20 European wild felids. All sequences showed 100% similarity to B. canis by BLAST analysis. Conversely, Cytb and COI analyses revealed the presence of two Babesia spp., B. pisicii n. sp., which we herein describe, and B. canis. The pairwise comparison of both mitochondrial genes of B. pisicii n. sp. showed a genetic distance of at least 10.3% from the most closely related species, B. rossi. Phylogenetic analyses of Cytb and COI genes revealed that B. pisicii n. sp. is related to the so-called “large” canid-associated Babesia species forming a separate subclade in a sister position to B. rossi.