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.