The global threat of neglected tropical diseases (NTDs) constitutes a public health issue in underdeveloped countries. Zoonotic helminthiases are the most common human NTD agents in developing countries in sub-Saharan Africa, Asia, and the Americas, causing a global burden of disease that exceeds that of more recognized infectious diseases such as malaria and tuberculosis. Wild canids are well-known mammals that act as natural reservoirs of zoonotic-relevant helminthiasis worldwide, thus playing a pivotal role in their epidemiology and transmission to humans. Here we evaluate the occurrence of zoonotic gastrointestinal helminths in two Neotropical wild canid species from the Amazonian and Andean regions of Colombia, i.e., the bush dog (Speothos venaticus) and the crab-eating fox (Cerdocyon thous). We recovered tapeworm proglottids from bush dog fecal samples and identified them molecularly as the canine-specific lineage of Dipylidium caninum by using cytochrome c oxidase subunit I (cox1) gene sequences. Moreover, examination of a crab-eating fox during necropsy revealed the presence of non-embryonated eggs of the neglected nematode Lagochilascaris cf. minor, in addition to eggs and gravid proglottids of the cestode Spirometra mansoni. These findings represent the first report of zoonotic-relevant cestodes, i.e., D. caninum ("canine genotype"), S. mansoni, and the nematode L. cf. minor, in bush dogs and crab-eating foxes as final hosts. The occurrence of these zoonotic helminthiases in wild canid species calls for regular monitoring programs to better understand the epidemiology and transmission routes of neglected dipylidiasis, lagochilascariosis, and sparganosis in South America.
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Species of Sarcocystis use various vertebrates as intermediate or definitive hosts in their life cycles. One of these is snakes, whose role as intermediate hosts for these protozoans is scarce; in fact, there are six records, but only three with molecular characterization. An imported green tree python was involved in the morphological and molecular characterization (four loci) of a new species of Sarcocystis localized in skeletal muscles. Sarcocystis moreliae sp. nov. has a type 1 sarcocyst with a smooth wall and is genetically similar (at the 18S rRNA gene) to two unnamed species of Sarcocystis found in Lytorhynchus diadema from Oman and Varanus salvator macromaculatus from Malaysia, but their detailed comparison is impossible. The new species showed lower similarity to its congeners in other loci (28S rRNA, ITS1, and cox1). This is the first morphological and genetic characterization of a Sarcocystis species in snakes of the genus Morelia, particularly M. viridis, using four loci, but more data are needed to fill the knowledge gap about snakes as intermediate hosts of Sarcocystis.
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Tick-transmitted apicomplexans of the genera Cytauxzoon and Hepatozoon affect a wide range of felids worldwide, but little is known about them. Recently, several studies addressed the species circulating in Europe, their distribution, and their hosts. Molecular assays are the method of choice for their detection. Unfortunately, conventional PCRs already described are time- and cost-consuming and specific for either Hepatozoon or Cytauxzoon detection. This study was developed to evaluate (i) the occurrence of Cytauxzoon and Hepatozoon in felids using a fast and cost-saving real-time PCR capable of detecting both protozoa simultaneously, (ii) the distribution of Cytauxzoon and Hepatozoon species in north-eastern Italy, and (iii) the involvement of other susceptible felid hosts in the same area. An SYBR® Green-based real-time PCR with primers targeting the 18S-rRNA was validated and applied to 237 felid samples, i.e., whole blood from 206 domestic cats and 12 captive exotic felids, and tissues from 19 wildcats. Positive results were obtained by melting temperature curve analysis due to the specific melting peak (i.e., 81°C Cytauxzoon spp.; 78-78.5°C Hepatozoon spp.). Positive samples were subjected to conventional PCR, followed by sequencing for species identification. Phylogenetic analyses were performed to assess relatedness among European isolates. Data on domestic cats (age class, sex, origin, management, and lifestyle) were recorded, and statistical analyses were performed to identify potential risk factors. A total of 31 (15%) domestic cats were positive for Hepatozoon spp. (i.e., 12 for H. felis, 19 for H. silvestris), while six (2.9%) for C. europaeus. The prevalence of Hepatozoon felis was significantly (p < 0.05) higher in domestic cats, while H. silvestris was higher in strays and animals from the Eastern region (i.e., Friuli-Venezia Giulia). Cytauxzoon europaeus was detected only in stray cats from Friuli-Venezia Giulia (province of Trieste). Among captive felids, one tiger was infected with H. felis and another with H. silvestris; eight out of 19 (42%) wildcats were positive for Hepatozoon spp. (i.e., six with H. felis, two with H. silvestris) and four out of 19 (21%) for Cytauxzoon europaeus. Outdoor lifestyle and origin (i.e., Friuli-Venezia Giulia region) were the most relevant risk factors for H. silvestris and C. europeus infections. Conversely, H. felis was most frequently isolated from domestic cats, suggesting different modes of transmission.
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Intervertebral disc extrusion (IVDE) is a common neurological condition in many dog breeds. This study aimed to describe this condition in Yorkshire terriers (YT) and calculate the prevalence of this condition amongst the YTs with neurological diseases. This is a double-centre retrospective study which was conducted in two arms. The first part of the study, describing the clinical features and prognosis of cervical (C) IVDE in YTs, is based on data from 2005 to 2021. The second part of the study calculated the prevalence of C IVDE amongst the YTs with neurological diseases based on data from 2016 to 2021. A retrospective search through the medical records was conducted. YTs with C IVDE diagnosed with MRI and confirmed surgically were eligible for inclusion in this study. Sixty YTs were included in the first part of the study. There were 48 (80%) dogs with acute onset and 12 (20%) with chronic onset with acute deterioration. Ambulation was preserved in 31 (51.7%) dogs on admission, and the remaining 29 (48.3%) dogs were non-ambulatory. No significant association was found between ambulation on admission and recovery status (p = 0.547). Seventy-three intervertebral spaces were treated during the surgical intervention. Relapses were seen in seven (11.7%) dogs. Forty-nine (81.7%) dogs were ambulatory at discharge. A complete recovery was observed in 46 (76.7%) dogs; the remaining dogs (14, 23.3%) were classified as incomplete recovery. A significant difference was found in time to ambulation (p = 0.0238) and time to discharge (p = 0.0139) between the on-admission ambulatory and non-ambulatory dogs. Three hundred and eight YTs were diagnosed with neurological diseases between 2016 and 2021 in one referral centre. C IVDE was diagnosed in 31 (10.06%) dogs. This is the first study explicitly describing the C IVDE in YTs and establishing the prevalence of this condition amongst YTs with other neurological disorders.
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Bovine papillomavirus type 1 L1 protein was produced in a baculovirus expression system and purified as virus-like particles (VLPs) by affinity chromatography using lectins. The morphological integrity of VLPs was confirmed by electron microscopy. Differences between the two detected variants were deciphered by mass spectrometry of peptides (MALDI-TOF). Mice were immunized with purified VLPs in doses of 10, 25, or 50 μg in combination with 1% saponin and 15% alhydrogel per dose as adjuvants. Analysis of the humoral immune response revealed increased levels of specific antibodies detected 3 weeks after the first immunization in all groups of animals. This was further significantly increased by the booster applied 3 weeks after the first dose, with the best immune response in a group of mice immunized by the largest dose of antigen. BPV1 L1 VLPs purified by affinity chromatography using lectins could be used for prophylactic immunization in veterinary medicine.
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