Molecular detection of Toxoplasma gondii in domestic and wild guinea pigs (Cavia spp.) from the Marangani district in Cuzco, Peru.

Toxoplasmosis is a foodborne disease caused by the protozoan Toxoplasma gondii, and transmitted to humans by eating raw or undercooked meat, mainly. Poultry, beef, and pork are the main meats consumed in Peru; despite this, guinea pig meat is also widely consumed. For this reason, the objective of this study was to molecularly detect T. gondii in domestic and wild guinea pigs from the Marangani district in Cuzco, Peru, and identify some risk factors associated with this pathogen. DNA was extracted from the brain tissue samples of guinea pigs (30 domestic and 30 wild), and PCR protocols were used to amplify the internal transcribed spacer (ITS-1) region and a 529 bp fragment from the T. gondii genome. T. gondii DNA was detected in 14 (23.3%) guinea pigs. T. gondii frequency was 33.3% in domestic guinea pigs and 13.3% in wild guinea pigs. Our results demonstrated that guinea pigs represent an important source for T. gondii infection in human populations in this locality.

Surveillance of SARS-CoV-2, rotavirus, norovirus genogroup II, and human adenovirus in wastewater as an epidemiological tool to anticipate outbreaks of COVID-19 and acute gastroenteritis in a city without a wastewater treatment plant in the Peruvian Highlands.

The coronavirus disease 2019 (COVID-19) pandemic has demonstrated that Wastewater Based Epidemiology is a fast and economical alternative for monitoring severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) at the community level in high-income countries. In the present study, wastewater from a city in the Peruvian Highlands, which lacks a wastewater treatment plant, was monitored for one year to assess the relationship between the concentration of SARS-CoV-2 and the reported cases of COVID-19 in the community. Additionally, we compared the relationship between rotavirus (RV), norovirus genogroup II (NoV GGII), and human adenovirus (HAdV) with the number of reported cases of acute gastroenteritis. Before commencing the analysis of the samples, the viral recovery efficacy of three processing methods was determined in spiked wastewater with SARS-CoV-2. This evaluation demonstrated the highest recovery rate with direct analysis (72.2 %), as compared to ultrafiltration (50.8 %) and skimmed milk flocculation (5.6 %). Wastewater monitoring revealed that 72 % (36/50) of the samples tested positive for SARS-CoV-2, with direct analysis yielding the highest detection frequency and quantification of SARS-CoV-2. Furthermore, a strong correlation was observed between the concentration of SARS-CoV-2 in wastewater and the reported cases of COVID-19, mainly when we shift the concentration of SARS-CoV-2 by two weeks, which allows us to anticipate the onset of the fourth and fifth waves of the pandemic in Peru up to two weeks in advance. All samples processed using the skimmed milk flocculation method tested positive and showed high concentrations of RV, NoV GGII, and HAdV. In fact, the highest RV concentrations were detected up to four weeks before outbreaks of acute gastroenteritis reported in children under four years of age. In conclusion, the results of this study suggest that periodic wastewater monitoring is an excellent epidemiological tool for surveillance and can anticipate outbreaks of infectious diseases, such as COVID-19, in low- and middle-income countries.

Seroprevalence of Toxoplasma gondii and associated risk factors in cats from Lima, Peru.

Toxoplasmosis is an important zoonotic disease worldwide caused by the intracellular parasite Toxoplasma gondii. Cats, being hosts, act as a good sentinel for this disease. Thus, the objective of this study was to determine the seroprevalence of T. gondii in cats by examining serum from seven districts of Metropolitan Lima, Peru. We collected and analyzed 303 cat serum samples, and the owners answered a questionnaire about age, gender, lifestyle (indoors only and outdoors access) and feeding (commercial and home food). The samples were tested using indirect hemagglutination (IHA) for detection of antibody IgM and IgG. Among the animals analyzed, we found 52 (17.2%, 95% CI: 13.1% - 21.9%) seropositive cats to T. gondii, of which one corresponded to acute infection (IgM) and 51 to chronic infections (IgG). In addition, the presence of anti-T. gondii antibodies was associated with feeding commercial food and living outdoors access. Our results concluded that T. gondii infections are common in cats with owners in Metropolitan Lima and will be useful to future epidemiological studies.

Coccidiosis in clinically asymptomatic alpaca (Vicugna pacos) crias from the Peruvian Andes.

Parasites of the genus Eimeria are involved in the neonatal diarrhea complex of alpaca (Vicugna pacos) crias, and infection by Eimeria is commonly known as coccidiosis. There are limited reports of these protozoa in clinically asymptomatic crias. In this study, fecal samples from 78 clinically asymptomatic alpaca crias were analyzed to evaluate the prevalence, parasitological load, and diversity of Eimeria species. This study was conducted in the Quenamari community located in the Peruvian Andes (Marangani, Cuzco) at 4500 m above sea level. All fecal samples were examined for parasites using the quantitative McMaster and modified Stoll techniques. Microscopic examination showed the presence of Eimeria oocysts in 68 out of the 78 samples (87.18%). Among the 78 samples we found E. lamae in 67 (85.90%), E. punoensis in 49 (62.82%), E. alpacae in 42 (53.85%), E. macusaniensis in 32 (41.03%), and E. ivitaensis in four (5.13%). Regarding parasitized crias, overall there was a mean parasitological load of 43,920 oocysts per gram of feces (OPG). Eimeria lamae had the highest parasitological load (mean 206,600 OPG). These findings could be due to environmental contamination with oocysts of different Eimeria species. Additional research is needed to determine if this burden of coccidiosis could produce subclinical impacts to the health of alpaca crias.

Abdominal dioctophymosis in a domestic cat from the Peruvian rainforest confirmed morphologically and molecularly.

A case of abdominal dioctophymosis in a domestic cat was found in San Juan Bautista district, the Peruvian rainforest, in the Loreto department of Peru. The pet went to a veterinary clinic for a routine ovariohysterectomy during which a large nematode was found in the abdominal cavity. The nematode was morphologically identified as an adult female of Dioctophyme sp. A few morphological parameters, such as the vagina distance from the anterior part and the egg size, were different than D. renale. Partial sequences of the cytochrome c oxidase subunit I (cox1) and the small subunit 18S ribosomal RNA genes were compared with the references from public sequence database and showed a genetic identifies of 89.25% and 99.65% with D. renale, respectively. This is the first mitochondrial molecular analysis of a Dioctophyme specimen from South America and the results showed up to 12.5% nucleotide sequence variation in cox 1 gene of D. renale.

Morphological and molecular evidence of Oslerus osleri (Nematoda: Filaroididae) in the Andean fox (Lycalopex culpaeus).

Oslerus osleri is a cosmopolitan filaroid nematode that parasitizes the respiratory system of domestic and wild canids. Natural infection by O. osleri is reported in the Andean fox (Lycalopex culpaeus) in this study. Nematodes, enclosed in small and compact fibrous nodules of 1 to 5 mm in diameter, were found on the surface of the trachea near the bronchial bifurcation on four Andean foxes during necropsy (one from Cuzco, Peru and three from Northwestern Patagonia in Argentina). The nematodes were identified as O. osleri by morphological and molecular methods. Ribosomal and mitochondrial DNA analyses were performed amplifying the second internal transcribed spacer region (ITS-2), the partial cytochrome c oxidase 1 (cox1), and the large subunit of nuclear ribosomal RNA (LSU rRNA) genes. Sequences of the ITS-2 and LSU rRNA had a genetic variation of 1.5% and 1.0%, respectively, with previous sequences of O. osleri registered in Genbank. This is the first amplification of the cox1 gene of O. osleri and demonstrated an identity of 92% to Perostrongylus falciformis (KY365437), and 90% to Angiostrongylus cantonensis (KY779735) and Angiostrongylus costaricensis (AP017675).

Prevalence and molecular characterization of Sarcoptes scabiei from vicuñas (Vicugna vicugna) from Southern Peruvian Andes.

Sarcoptic mange is a disease caused by an infectious parasite in the vicuñas (Vicugna vicugna) from South America. Although molecular studies have provided much information about the epidemiology of this disease, this information is still unknown in vicuñas. This study determined the prevalence and molecular characterization of Sarcoptes scabiei from vicuñas from Southern Peruvian Andes. During the 2018 shearing season, 181 vicuñas were clinically evaluated for lesions compatible with mange. Sarcoptes scabiei was detected in 35 (19.3%) vicuñas, and 50 mites from 25 vicuñas were selected for molecular analyses of the mitochondrial (cox1) and nuclear (ITS2) genetic markers. Molecular analyses of the cox1 and ITS2 sequences showed an identity of 94­99% and 99.8­100% with previous S. scabiei sequences registered in the GenBank, respectively. Sequence polymorphisms were more evident in the ITS2 than in the cox1, but only the cox1 had an association with the host. Phylogenetic analysis of S. scabiei cox1 sequences from vicuñas showed a cluster with S. scabiei cox1 sequences from canids, suggesting that the origin of S. scabiei from vicuña is associated with canid mites. This research is the first molecular analysis of S. scabiei from vicuñas. Future molecular studies will be necessary to determine the species variety, geographic segregation and host­parasite adaptation for this vicuña's mite.

Cryptosporidium parvum as a risk factor of diarrhea occurrence in neonatal alpacas in Peru.

Cryptosporidiosis has been reported as an important cause of neonatal diarrhea and mortality in cattle, sheep, and other ruminants, but its impact on alpaca health has not been studied thoroughly. In this study, we have determined the prevalence and evaluated the role of cryptosporidiosis as a risk factor for diarrhea occurrence in newborn alpacas. During the calving season (January-March) of 2006, stool specimens (N = 1312) were collected from 24 herds of newborn alpacas in Puno and Cuzco, departments that account for the largest populations of alpacas in Peru. All the specimens were microscopically screened for Cryptosporidium spp. using the acid-fast technique. The association between Cryptosporidium detection and diarrhea was analyzed using χ2 test and generalized lineal model. Cryptosporidium species were determined by PCR-RFLP analysis of the small subunit rRNA gene. Cryptosporidium oocysts were detected in 159 of 1312 (12.4%) newborn alpacas. Results of the analyses demonstrated that crypstosporidiosis was significantly associated with diarrhea (PR = 3.84; CI95% 2.54-5.81; p < 0.0001). Only Cryptosporidium parvum was detected in the 153 Cryptosporidium-infected animals. Thus, there is an association of C. parvum infection with diarrhea in neonatal alpacas.

Alopecia a potential adverse side effect of albendazole use in alpacas.

Albendazole is a benzimidazole derivative with anthelmintic activity. It is the treatment of choice for fasciolosis. The use of albendazole in South American camelids is common, however, there are no studies about the pharmacokinetics and pharmacodynamics of albendazole in alpacas and llamas. In the present study, a case of fiber loss (alopecia) in alpacas is described because of the suspected use of a high dose of albendazole. In a fasciolosis control program of an alpaca ranch located in the district of Nuñoa in Puno, Peru, 2184 alpacas were oral treated with albendazole (35-40 mg/kg). After 2 weeks of treatment the alpacas began to show loss of fiber in the abdomen, flanks and neck. The alpacas showed no other sign of disease. The alpacas recovered their fiber after 6 months. We suggest studies are needed to determine the safe dose of albendazole in alpacas.

Infección natural por Fasciola hepatica en cérvidos del Perú / Natural infection of Fasciola hepatica in cervids from Peru

En el presente trabajo se registra la infección natural por Fasciola hepatica en un venado de cola blanca (Odocoileus virginianus) y en una taruca (Hippocamelus antisensis), ambos procedentes del departamento de Cusco. Los animales fueron remitidos al Instituto Veterinario (IVITA-Maranganí, FMV, UNMSM) por las autoridades del Servicio Nacional de Flora y Fauna (SERFOR, Sede Cusco). Durante la necropsia de los animales se colectaron seis trematodos de los conductos biliares, los cuales fueron preservados en etanol al 70%. Las observaciones morfológicas indicaron que se trataban de F. hepatica. Esto fue confirmado analizando el ADN mitocondrial de los parásitos amplificando parcialmente los genes citocromo c oxidasa subunidad 1 (cox1) y el NADH deshidrogenasa subunidad 1 (nad1). El análisis de estos genes tuvo una identidad mayor al 99% comparado con registros del banco de genes (GenBank). El presente estudio demuestra la presencia de F. hepatica en estos cérvidos, agregando así dos nuevos hospederos definitivos para el parásito.

Natural infection by Fasciola hepatica is recorded in a white-tailed deer (Odocoileus virginianus) and a taruca (Hippocamelus antisensis), both from the department of Cusco. Animals were remitted to the Veterinary Institute (IVITA-Maranganí, FMV, UNMSM) by the authorities of the National Service of Flora and Fauna (SERFOR, Cusco Headquarters). Six trematodes were collected from the bile ducts during the necropsy of the animals, and they were preserved in 70% ethanol. Morphological analysis indicated that they correspond to F. hepatica. This was confirmed by analyzing of the mitochondrial DNA of the parasites by partially amplifying the cytochrome c oxidase subunit 1 (cox1) and the NADH dehydrogenase subunit 1 (nad1) genes. Analysis of these genes had an identity greater than 99% compared to genes from GenBank. The present study demonstrates the occurrence of F. hepatica in these cervids, thus adding two new definitive hosts for the parasite.

Morphological and molecular identification of Spirocerca lupi (Nematoda: Spiruridae) found in the Andean fox (Lycalopex culpaeus).

Lesions compatible with spirocercosis were found in the esophagus and aorta of an Andean fox from Cuzco, Peru. The esophageal and aortic lesions were 5.5 and 1.5 cm in diameter, respectively. A total of 12 adult nematodes (6 males and 6 females) were collected from the esophageal lesion, and all were identified as Spirocerca lupi by morphological and molecular methods. Molecular characterization was performed by analyzing two sources of the cox1 gene, and the sequences were compared with previous S. lupi sequences from other work deposited in GenBank. Analysis of the partial cox1 gene from S. lupi (n = 3) showed 2 haplotypes and had 95-99% nucleotide similarity to previously described sequences. Also, molecular analysis showed that S. lupi is a very diverse group, due to the genetic variability of the partial sequences of the cox1 gene of Spirocerca. This study is the first to report finding of spirocercosis in the Andean fox.

The alpaca (Vicugna pacos) as a natural intermediate host of Taenia omissa (Cestoda: Taeniidae).

Three metacestodes were collected from the mesentery and the surface of the liver of three adult alpacas (Vicugna pacos) in a slaughterhouse located in Puno, Peru. Various features of the metacestodes were observed for morphological identification. A molecular diagnosis was performed by PCR-based sequencing of mitochondrial genes of cytochrome c oxidase subunit 1 (cox1) and the NADH dehydrogenase subunit 1 (nad1). All metacestodes were identified as Taenia omissa by morphology and molecular methods The isolates from alpacas showed significant sequence similarity with previously reported isolates of T. omissa (95.7-98.1% in cox1 and 94.6-95.1% in nad1). Our report is the first to detect T. omissa metacestodes in alpacas and to reveal that alpacas are natural intermediate hosts for this parasite.

First finding of nymphal stages of Linguatula serrata in a South American camelid, a vicuña from Peru.

Linguatula serrata, a pentastomid, was found parasitizing the lungs of a vicuña (Vicugna vicugna) from Cuzco, Peru. A total of 13 larvae were found encysted in the parenchymal tissue of the lungs. All larvae were identified as nymphal stages of L. serrata by morphological methods Diagnosis was confirmed by molecular analysis amplifying the cytochrome c oxidase 1 gene of three nymphs. Nucleotide sequences from the isolates were compared to previous sequences from GenBank, and it showed high similarity between them (>99%). This finding constitutes the first detection of L. serrata in a South American camelid.

Evaluation of activity of triclabendazole against Taenia solium metacestode in naturally infected pigs.

OBJECTIVE: To assess the efficacy of triclabendazole (TCBZ) in porcine cysticercosis. METHODS: Eighteen naturally infected cysticercosis pigs were divided into 3 groups of 6 individuals each. The first group was treated orally with TCBZ at a single dose of 30 mg/kg of body weight, the second group was treated orally with oxfendazole at a single dose of 30 mg/kg of body weight and the third group received a placebo (control group). All animals were kept under the same management conditions. The pigs were euthanized 17 wk post-treatment and the number of surviving cysts in muscles was assessed and compared between groups. RESULTS: All pigs treated with oxfendazole had only degenerated cysts in their carcasses. In contrast, TCBZ had very little effect against the parasitic cysts. Cysts from pigs in the TCBZ group looked apparently normal after treatment. However, histological evaluation showed a mild to moderate degree of inflammation. CONCLUSIONS: TCBZ is not an efficacious drug against Taenia solium cysticercosis in swine using a single dose.

Finding of pentastomes of genus Reighardia (Pentastomida) in the Belcher's gull (Larus belcheri).

This report describes the finding of Reighardia sp. (Pentastomida) infecting the air sac of two Belcher's gulls (Larus belcheri) found dead on the beaches of Pucusana, a district in southern Lima, Peru. Three pentastomes were collected from two Belcher's gulls. Then, they were morphologically and molecular analyzed. Molecular characterization of the parasite was achieved by amplifying a fragment of the small subunit ribosomal RNA gene (SSU rRNA). Based on both morphological and molecular data the pentastomes were identified as pentastomes of the genus Reighardia. This is the first report showing that the Belcher's gull is a new natural definitive host for this pentastome.

The taruca (Hippocamelus antisensis) and the red brocket deer (Mazama americana) as intermediate hosts of Taenia hydatigena in Peru, morphological and molecular evidence.

In the present report metacestodes were collected from the mesentery of a taruca (Hippocamelus antisensis) and from the omentum of a red brocket deer (Mazama americana) in Peru. Various metacestodes parameters, including rostellar hook characteristics, were measured. Molecular analysis was performed to amplify the mitochondrial cytochrome c oxidase subunit 1 gene from metacestode isolates. Metacestodes were identified as T. hydatigena by morphology and molecular methods. This constitutes the first molecular detection of T. hydatigena metacestodes in the taruca and the red brocket deer and demonstrates that these animal species are natural intermediate hosts for this parasite.

New insights in cysticercosis transmission.

Taenia solium infection causes severe neurological disease in humans. Even though infection and exposure to swine cysticercosis is scattered throughout endemic villages, location of the tapeworm only explains some of the nearby infections and is not related to location of seropositive pigs. Other players might be involved in cysticercosis transmission. In this study we hypothesize that pigs that carry nematodes specific to dung beetles are associated with cysticercosis infection and/or exposure. We carried out a cross-sectional study of six villages in an endemic region in northern Peru. We euthanized all pigs (326) in the villages and performed necropsies to diagnose cysticercosis. For each pig, we counted cysticerci; measured anti-cysticercus antibodies; identified intestinal nematodes; tabulated distance to nearest human tapeworm infection; and recorded age, sex, productive stage, and geographic reference. For the purpose of this paper, we defined cysticercosis infection as the presence of at least one cysticercus in pig muscles, and cysticercosis exposure as seropositivity to anti-cysticercus antibodies with the presence of 0-5 cysticerci. Compared to pigs without nematode infections, those pigs infected with the nematode Ascarops strongylina were significantly associated with the presence of cysticerci (OR: 4.30, 95%CI: 1.83-10.09). Similarly, pigs infected with the nematode Physocephalus sexalatus were more likely to have cysticercosis exposure (OR: 2.21, 95%CI: 1.50-3.28). In conclusion, our results suggest that there appears to be a strong positive association between the presence of nematodes and both cysticercosis infection and exposure in pigs. The role of dung beetles in cysticercosis dynamics should be further investigated.

Occurrence of Giardia duodenalis assemblages in alpacas in the Andean region.

In this study, 352 fecal samples were analyzed for G. duodenalis from alpaca mothers and crias from three different areas of highland in Peru. The triosephosphate isomerase (TPI) gene of Giardia was amplified using a nested PCR protocol. Forty-six G. duodenalis-PCR positive samples were sequenced. G. duodenalis assemblage A was the most frequent followed by assemblage E. The former was seen in 37 animals whereas the latter was seen in nine. Most of the assemblage A infections were caused by the A1 subtype of sub-assemblage AI, except for three, which were caused by the A2 subtype of sub-assemblage AI. Assemblage A was found in all three geographic regions, while assemblage E was detected in crias from two regions. Among the four alpaca mothers positive for Giardia, three had assemblage AI and one had assemblage AII. Results of this study indicate that possible zoonotic transmission human to alpacas.

Trombiculiasis caused by chigger mites Eutrombicula (Acari: Trombiculidae) in Peruvian alpacas.

Trombiculiasis is an infestation caused by larvae members of the family Trombiculidae, common called chigger mites. In this study is presented the first case of trombiculiasis caused by the infestation of chigger mite Eutrombicula in alpacas from Peru. Twenty-two alpacas of a total of 130 animals were infested by Eutrombicula sp. The chigger mite location was only in the face skin folds and around the eyes. In addition, all alpacas infested had alopecia and dermatitis in the infected zone.

Efficacy of a single oral dose of oxfendazole against Fasciola hepatica in naturally infected sheep.

The efficacy of a single oral dose of 30 mg/kg of oxfendazole against Fasciola hepatica was evaluated in a controlled study in naturally infected sheep. Sheep were diagnosed by stool microscopy after sedimentation, and positive animals were randomized to oxfendazole (N = 20) or no treatment (N = 20). A new stool exam was performed 10 days after treatment. All stool microscopies were performed masked to the treatment group. No side effects were noticed. All sheep in the control group remained infected with similar counts of eggs per gram of stools. None of the animals in the treatment group showed Fasciola eggs in stools after 10 days of treatment. A single dose of oxfendazole is highly effective against F. hepatica, providing a new drug alternative for the control of fascioliasis or integrated zoonosis control.