Canine parasitoses in north Patagonia (Argentina): comparison between different social and environmental factors.

In many countries, urban areas are commonly contaminated with canine feces that spread parasites, a situation related to the natural and social environment. Dogs having free access to public areas therefore constitutes a health risk. Because in Patagonia there are no records of city comparisons that consider the number of free-roaming dogs and the local environmental and socioeconomic characteristics, in this study we analyse and compare canine parasitoses in two cities of Rio Negro province, Argentina. Canine feces were collected from public areas of El Bolsón and Cipolletti cities and examined using enrichment methods, Kinyoun stain and immunoassay (Copro-Elisa). The total percentage of positive feces in El Bolsón was 68.95% and 16 parasite taxa being identified, whereas in Cipolletti the total positive feces was 41.1%, with 11 taxa. Both cities presented more helminth species than protozoans. The variables that explained the occurrence of all parasites were rainfall and socioeconomic stratum with the highest values being found in low-income areas.Soil moisture is conditioned by rainfall. This study highlights the presence of the zoonotic Echinococcus sp. in dogs in urban areas of the Rio Negro province. The occurrence of this parasite in Cipolletti is unexpected, given that this city is not considered an endemic zone for this disease, which is not included in local monitoring and control plans.

Presence of Echinococcus granulosussensulato in the endoparasitic fauna of feral dogs in Tierra del Fuego, Argentina.

The feral dog population from Tierra del Fuego (Argentina) has affected provincial sheep production, and is responsible for the decrease of 43% of the sheep stock. In addition, feral dogs preys on the native fauna, especially the guanaco (Lama guanicoe), a species protected by law. Except for some data on the aforementioned impact, very little is known about feral dogs features in this region, such as distribution, abundance, diet, reproduction, health status, among others. Therefore, a descriptive observational study was carried out in the period 2018-2021 in Tierra del Fuego, with the aim to recognize the endoparasitic fauna of these dogs, with emphasis on Echinococcus granulosus sensu lato (s.l.). Feces samples were collected from 83 feral dogs captured in 15 farms, which were evaluated using flotation and sedimentation coproparasitological techniques, and optical microscope observation. These qualitative methods showed that 66.3% of the samples were positive. Nine parasitic forms were found, including eggs of taenids, Toxocara canis and Toxascaris leonina, and Sarcocystis spp. oocysts and Giardia spp. cysts. Samples with more than one parasitic form were also observed, among which the Taenia/Sarcocystis spp. association was the most frequent. For the diagnosis of canine echinococcosis, the detection of Echinococcus spp. antigens by Copro-ELISA and the molecular confirmation by Copro-PCR were performed. The Copro-ELISA test was positive in 10/80 (12.5%) of the samples processed, while 11 samples (13.8%) were positive by Copro-PCR. Their distribution showed that 9 (81.8%) corresponded to farms with sheep farming, and 2 (18.2%) with cattle farming. The presence of E. granulosus s.l. in feral dogs from Tierra del Fuego is the most relevant finding in this study. These results confirm that feral dogs have access to viscera infected with hydatid cysts. However, the intermediate hosts involved (sheep, cattle or guanacos) should be investigated. Its impact on the transmission of cystic echinococcosis in this region should be evaluated, as well as the adequacy of the disease prevention and control actions implemented in Tierra del Fuego should be considered. Finally, the existence of a wild cycle of the disease should be evaluated, in which feral dogs would act as a source of infection for other animals and humans.

Botulismo del lactante: estudio descriptivo en una unidad de cuidados intensivos pediátricos / Infant botulism: a descriptive study in a pediatric intensive care unit

Introducción. El botulismo del lactante (BL) es la forma más frecuente de botulismo humano en Argentina. El objetivo es describir aspectos esenciales del diagnóstico y tratamiento de pacientes con BL internados en el servicio de terapia intensiva pediátrica (STIP). Métodos. Estudio observacional, descriptivo y retrospectivo. Se utilizó la base de datos del STIP con diagnóstico de BL en el período 2005-2020. Se registraron variables demográficas, métodos de diagnóstico, días de asistencia respiratoria mecánica convencional (ARMC), de ventilación no invasiva (VNI), estadía en STIP, mortalidad al alta hospitalaria. Resultados. Se registraron 21 pacientes con BL; 14 pacientes fueron varones, con una mediana de edad de 5 meses (RIC 2-6 m). El diagnóstico se realizó mediante técnica de bioensayo y se detectó la toxina en suero en 12 pacientes. Uno solo no requirió ARMC; 1 paciente fue traqueostomizado; 18 pacientes recibieron antibióticos; 5 recibieron VNI. Ningún paciente recibió antitoxina y no hubo fallecidos. La mediana de estadía hospitalaria fue 66 días (RI: 42-76); de internación en STIP, 48 días (RI: 29-78); y de ARMC, 37 días (RI: 26-64). La demora en la confirmación diagnóstica fue 15,8 ± 4,8 días. Conclusiones. La totalidad de los pacientes fueron diagnosticados con la técnica de bioensayo, que generó un tiempo de demora diagnóstica que excede los lapsos recomendados para la administración del tratamiento específico. Ningún paciente recibió tratamiento específico. El BL presentó baja mortalidad, pero tiempos de ARM e internación prolongados, que se asocian a infecciones sobreagregadas y uso frecuente de antibióticos.

Introduction. Infant botulism (IB) is the most common form of human botulism in Argentina. Our objective was to describe the main aspects of diagnosis and management of patients with IB admitted to the pediatric intensive care unit (PICU). Methods. Observational, descriptive, and retrospective study. The PICU database with IB diagnosis in 2005­2020 period was used. Demographic variables, diagnostic methods, days of conventional mechanical ventilation (CMV), non-invasive ventilation (NIV), length of stay in the PICU and mortality upon hospital discharge were recorded. Results. In total, 21 patients with IB were recorded; 14 were male, their median age was 5 months (IQR: 2­6 m). Diagnosis was made by bioassay, and the toxin was identified in the serum of 12 patients. Only 1 patient did not require CMV; 1 patient had a tracheostomy; 18 patients received antibiotics; 5 received NIV. No patient was administered antitoxin and no patient died. The median length of stay in the hospital was 66 days (IQR: 42­76); in the PICU, 48 days (IQR: 29­78); and the median use of CMV, 37 days (IQR: 26­64). The delay until diagnostic confirmation was 15.8 ± 4.8 days. Conclusions. All patients were diagnosed using the bioassay technique, which resulted in a diagnostic delay that exceeds the recommended period for the administration of a specific treatment. No patient received a specific treatment. IB was related to a low mortality, but also to prolonged use of MV and length of hospital stay, which were associated with cross infections and frequent antibiotic use.

Differences in clinical aspects of human cystic echinococcosis caused by Echinococcus granulosus sensu stricto and the G6 genotype in Neuquén, Argentina.

Most human cystic echinococcosis (CE) cases worldwide are attributed to Echinococcus granulosus sensu stricto (s.s), followed by the G6 and G7 genotypes. While E. granulosus s.s. has a cosmopolitan distribution, the G6 genotype is restricted to areas where camels and goats are present. Goats are the primary livestock in the Neuquén province in Argentina where the G6 genotype has been reported to be responsible for a significant percentage of CE human cysts genotyped. In the present study, we genotyped 124 Echinococcus cysts infecting 90 CE-confirmed patients. Echinococcus granulosus s.s. was identified in 51 patients (56.7%) with 81 cysts and the G6 genotype in 39 patients (43.3%) harbouring 43 cysts. Most CE cases ≤18 years were male suggesting pastoral work could be a risk factor for the infection. Echinococcus granulosus s.s. was significantly found more frequently in the liver (32/51 patients) and the G6 genotype in the lungs and extrahepatic localizations (27/39). The patients infected with E. granulosus s.s., presented up to 6 cysts while patients infected with G6 presented a maximum of 2. The diameter of lung cysts attributed to E. granulosus s.s. was significantly larger compared to lung cysts from G6. Following the WHO ultrasound classification of liver cysts, we observed inactive cysts in 55.6% of G6 cysts and only 15.3% of E. granulosus s.s cysts. In conclusion, we provide evidence of differences in clinical aspects of CE caused by E. granulosus s.s. and the G6 genotype of E. granulosus s.l. complex infecting humans.

Infant botulism: a descriptive study in a pediatric intensive care unit. / Botulismo del lactante: estudio descriptivo en una unidad de cuidados intensivos pediátricos.

Introduction. Infant botulism (IB) is the most common form of human botulism in Argentina. Our objective was to describe the main aspects of diagnosis and management of patients with IB admitted to the pediatric intensive care unit (PICU). Methods. Observational, descriptive, and retrospective study. The PICU database with IB diagnosis in 2005-2020 period was used. Demographic variables, diagnostic methods, days of conventional mechanical ventilation (CMV), non-invasive ventilation (NIV), length of stay in the PICU and mortality upon hospital discharge were recorded. Results. In total, 21 patients with IB were recorded; 14 were male, their median age was 5 months (IQR: 2-6 m). Diagnosis was made by bioassay, and the toxin was identified in the serum of 12 patients. Only 1 patient did not require CMV; 1 patient had a tracheostomy; 18 patients received antibiotics; 5 received NIV. No patient was administered antitoxin and no patient died. The median length of stay in the hospital was 66 days (IQR: 42-76); in the PICU, 48 days (IQR: 29-78); and the median use of CMV, 37 days (IQR: 26-64). The delay until diagnostic confirmation was 15.8 ± 4.8 days. Conclusions. All patients were diagnosed using the bioassay technique, which resulted in a diagnostic delay that exceeds the recommended period for the administration of a specific treatment. No patient received a specific treatment. IB was related to a low mortality, but also to prolonged use o  MV and length of hospital stay, which were associated with cross infections and frequent antibiotic use.

Introducción. El botulismo del lactante (BL) es la forma más frecuente de botulismo humano en Argentina. El objetivo es describir aspectos esenciales del diagnóstico y tratamiento de pacientes con BL internados en el servicio de terapia intensiva pediátrica (STIP). Métodos. Estudio observacional, descriptivo y retrospectivo. Se utilizó la base de datos del STIP con diagnóstico de BL en el período 2005-2020. Se registraron variables demográficas, métodos de diagnóstico, días de asistencia respiratoria mecánica convencional (ARMC), de ventilación no invasiva (VNI), estadía en STIP, mortalidad al alta hospitalaria. Resultados. Se registraron 21 pacientes con BL; 14 pacientes fueron varones, con una mediana de edad de 5 meses (RIC 2-6 m). El diagnóstico se realizó mediante técnica de bioensayo y se detectó la toxina en suero en 12 pacientes. Uno solo no requirió ARMC; 1 paciente fue traqueostomizado; 18 pacientes recibieron antibióticos; 5 recibieron VNI. Ningún paciente recibió antitoxina y no hubo fallecidos. La mediana de estadía hospitalaria fue 66 días (RI: 42-76); de internación en STIP, 48 días (RI: 29-78); y de ARMC, 37 días (RI: 26-64). La demora en la confirmación diagnóstica fue 15,8 ± 4,8 días. Conclusiones. La totalidad de los pacientes fueron diagnosticados con la técnica de bioensayo, que generó un tiempo de demora diagnóstica que excede los lapsos recomendados para la administración del tratamiento específico. Ningún paciente recibió tratamiento específico. El BL presentó baja mortalidad, pero tiempos de ARM e internación prolongados, que se asocian a infecciones sobreagregadas y uso frecuente de antibióticos.

A cross-sectional study of free-roaming dogs in a Patagonian city: Their distribution and intestinal helminths in relation to socioeconomic aspects of neighborhoods.

Human and dog health are inextricably linked, and although our relationship with dogs brings numerous benefits for our well-being, it is known that they can transmit disease. Bariloche is a Patagonian tourist city with heterogeneous social composition. This cross-sectional study evaluates the population of free-roaming dogs and their intestinal parasites in relation to the socioeconomic level of the city's human population. Census areas were used as survey units, stratified in three levels according to socioeconomic status. The free-roaming dog population was estimated by walking around each census area. Eight fresh feces per census count area were collected and analyzed using coproparasitological flotation tests, and CoproELISA was used to detect Echinococcus sp. A total of 858 free-roaming dogs were registered along 40.9 km, with significant differences between socioeconomic strata: the highest numbers were found in the lowest income strata. Of the feces collected, 39.2% tested positive for parasites, those associated with a lower socioeconomic level having higher percentages of positive feces and a greater number of species. Eight species of helminths were found, some of which were zoonotic, such as Echinococcus sp., Toxocara canis, and Dibothriocephalus latus. The presence of parasites can be explained by the number of free-roaming dogs per census count area. The free-roaming dogs generally have owners, and their parasitic infection is strongly associated with the socioeconomic level of the population. The main problem is irresponsible pet care, which generates healthy conditions for both dogs and humans. Thus, both dogs and humans deserve effective ethical public policies.

Analysis of nad2 and nad5 enables reliable identification of genotypes G6 and G7 within the species complex Echinococcus granulosus sensu lato.

The larval stages of tapeworms in the species complex Echinococcus granulosus sensu lato cause a zoonotic disease known as cystic echinococcosis (CE). Within this species complex, genotypes G6 and G7 are among the most common genotypes associated with human CE cases worldwide. However, our understanding of ecology, biology and epidemiology of G6 and G7 is still limited. An essential first step towards this goal is correct genotype identification, but distinguishing genotypes G6 and G7 has been challenging. A recent analysis based on complete mitogenome data revealed that the conventional sequencing of the cox1 (366 bp) gene fragment mistakenly classified a subset of G7 samples as G6. On the other hand, sequencing complete mitogenomes is not practical if only genotype or haplogroup identification is needed. Therefore, a simpler and less costly method is required to distinguish genotypes G6 and G7. We compared 93 complete mitogenomes of G6 and G7 from a wide geographical range and demonstrate that a combination of nad2 (714 bp) and nad5 (680 bp) gene fragments would be the best option to distinguish G6 and G7. Moreover, this method allows assignment of G7 samples into haplogroups G7a and G7b. However, due to very high genetic variability of G6 and G7, we suggest to construct a phylogenetic network based on the nad2 and nad5 sequences in order to be absolutely sure in genotype assignment. For this we provide a reference dataset of 93 concatenated nad2 and nad5 sequences (1394 bp in total) containing representatives of G6 and G7 (and haplogroups G7a and G7b), which can be used for the reconstruction of phylogenetic networks.

The benefits of analysing complete mitochondrial genomes: Deep insights into the phylogeny and population structure of Echinococcus granulosus sensu lato genotypes G6 and G7.

Cystic echinococcosis (CE) is a zoonotic disease caused by the larval stage of the species complex Echinococcus granulosus sensu lato. Within this complex, genotypes G6 and G7 have been frequently associated with human CE worldwide. Previous studies exploring the genetic variability and phylogeography of genotypes G6 and G7 have been based on relatively short mtDNA sequences, and the resolution of these studies has often been low. Moreover, using short sequences, the distinction between G6 and G7 has in some cases remained challenging. The aim here was to sequence complete mitochondrial genomes (mitogenomes) to obtain deeper insight into the genetic diversity, phylogeny and population structure of genotypes G6 and G7. We sequenced complete mitogenomes of 94 samples collected from 15 different countries worldwide. The results demonstrated that (i) genotypes G6 and G7 can be clearly distinguished when mitogenome sequences are used; (ii) G7 is represented by two major haplogroups, G7a and G7b, the latter being specific to islands of Corsica and Sardinia; (iii) intensive animal trade, but also geographical isolation, have likely had the largest impact on shaping the genetic structure and distribution of genotypes G6 and G7. In addition, we found phylogenetically highly divergent haplotype from Mongolia (Gmon), which had a higher affinity to G6.

Genetic characterization of human hydatid cysts shows coinfection by Echinococcus canadensis G7 and Echinococcus granulosus sensu stricto G1 in Argentina.

Human cystic echinococcosis caused by the larval stage of Echinococcus granulosus sensu lato (s.l.) is a highly endemic disease in the province of Neuquén, Patagonia, Argentina. Human infections with E. granulosus sensu stricto (s.s.) G1 and Echinococcus canadensis G6 were reported in Neuquén in previous studies, whereas four genotypes were identified in livestock: G1, G3, G6, and G7. The aim of this study was to identify the genotypes of E. granulosus s.l. isolates from humans of Neuquén province, Patagonia, Argentina, through the 2005-2014 period. Twenty six hydatid cysts were obtained from 21 patients. The most frequent locations were the liver and lungs. Single cysts were observed in 81.0% of patients, and combined infection of liver and lungs was detected in 9.5% of cases. Partial sequencing of mitochondrial cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 1 (nad1) genes identified the presence of E. granulosus s.s. G1 (n = 11; 42.3%) including three different partial sequences; E. canadensis G6 (n = 14; 53.8%) and E. canadensis G7 (n = 1; 3.9%). Coinfection with G1 and G7 genotypes was detected in one patient who harbored three liver cysts. Most of the liver cysts corresponded to G1 and G6 genotypes. This study presents the first report in the Americas of a human infection with E. canadensis G7 and the second worldwide report of a coinfection with two different species and genotypes of E. granulosus s.l in humans. The molecular diversity of this parasite should be considered to redesign or improve the control program strategies in endemic regions.

First study about the development of adult Echinococcus canadensis G6 genotype of goat origin in experimentally infected dogs.

Echinococcus granulosus sensu lato (E. granulosus sl) must be considered as a species complex, comprising Echinococcus granulosus sensu stricto (E. granulosus ss, genotypes G1-G3), Echinococcus equinus (G4), Echinococcus ortleppi (G5) and Echinococcus canadensis (G6-G10) although the species status of E. canadensis is still controversial. These genotypes closely match the intermediate hosts associated strains described in earlier times among which E. canadensis G6 corresponds to the camel strain. As there are no studies concerning the development of adult stages of the G6 genotype from non-camel origin, the aims of the present study were: to characterize for the first time the development of E. canadensis G6 in dogs experimentally infected with protoscoleces derived from goats, to describe the resultant adult morphology, to evaluate the growth of their rostellar hooks from larval to adult stages and to determine the prepatent period of the strobilar stage of E. canadensis G6 derived from goats. The development of the strobilar stage of E. canadensis G6 genotype of goat origin was examined by studying the growth (variation of the total worm length) and segmentation in experimentally infected dogs at 14, 25, 35 and 56days post infection. A morphological characterization of 35-day-old worms as well as of larval and adult rostellar hooks was also carried out by conventional optical microscopic observations and/or by scanning electron microscopy. The prepatent period of the strobilar stage was assessed by microscopic examination of faeces from 2 infected dogs. Our results were compared with published data from the camel and other strains. The roles of the host, genotype and species in morphological and developmental features as well as the taxonomic position of E. canadensis G6 were discussed. The prepatent period of E. canadensis G6 genotype of goat origin was determined as at least, 41days. The obtained results contribute to increase the knowledge about the biology and genetics of E. granulosus sl complex and are also of practical usefulness for the design of disease control strategies.

A wide diversity of zoonotic intestinal parasites infects urban and rural dogs in Neuquén, Patagonia, Argentina.

The presence of parasites was investigated by the examination of 1944 dog faecal samples collected from urban (n=646) and rural (n=1298) areas of the province of Neuquén, Patagonia, Argentina. Parasitic agents (PA) were found in 37.86% of samples. A total of 15 different PA were detected, including Toxocara canis (16.35%), Taenia spp./Echinococcus spp. (12.65%), Trichurisvulpis (6.06%), Giardia spp. (1.29%), Toxascaris leonina (0.56%), Ancylostomacaninum (0.41%), Dipylidium caninum (0.31%), Diphyllobothrium spp. (0.10%), among others. Several of these PA are recognized as zoonotic agents. Therefore, the results of this investigation revealed that local population is exposed to a broad spectrum of zoonotic parasites by means of environmental contamination with dog faeces. Prevalence of PA was slightly higher in rural (40.06%) than in urban (33.44%) locations. Distribution of groups of PA (cestodes, nematodes, and protozoa) showed statistical differences between both habitats. Prevalence of cestodes (18.18%) and protozoa (11.86%) was significantly higher in the rural environment than in urban areas and nematodes (29.10%) were more frequent in urban locations. Infection of dogs with Linguatula serrata and Cryptosporidium sp. was demonstrated for the first time in Neuquén. Rural dogs of the study area are under hydatic disease control program, which includes treatment with praziquantel every 6 weeks; thus, the finding of high level of cestode infection in these areas is of great relevance. The epidemiology of zoonotic parasitic infections in urban and rural dogs showed different patterns and, in consequence, different control measurements should be applied in each location.

Heterogeneous distribution of human cystic echinococcosis after a long-term control program in Neuquén, Patagonia Argentina.

The present study is the first comprehensive analysis of human cystic echinococcosis (CE) epidemiological data carried out in the province of Neuquén, Patagonia Argentina, after 34 years of uninterrupted control program. A retrospective study of all the official records of confirmed human CE cases between 1995 and 2004 was carried out. A total of 1107 cases were reported. The overall mean annual incidence (MAI) was 24.4 per 100,000 inhabitants for the total population and 9.7/100,000 in the 0-14 year group. Distribution of cases by age and sex as well as frequency of cyst locations was analysed. Children accounted for 13.3% of total cases, indicating active transmission of the disease. Territorial distribution of cases was highly heterogeneous: MAI per 100,000 ranged from 7.9 in the Metropolitan Sanitary Area to 78.4 in the western rural areas. Higher values were recorded in small communities as Pilo Lil (800/100,000). MAI showed a significant decrease from 1995 to 1999 (43.9-18.8) but remains stable from 2000 to 2004 (15.9-15.5). These results suggest that standard control measures, despite long-term implementation, are not able to produce a sustained improvement of the epidemiological status of the disease. Further studies about local transmission cycles, definite and intermediate hosts present, Echinococcus granulosus strains or cultural behaviours in small communities are required in order to adequate the control actions in Neuquén.

Parasitosis intestinales y su relación con factores socioeconómicos y condiciones de habitat en niños de Neuquén, Patagonia, Argentina / Intestinal parasitosis in relation to socioeconomic factors and habitat conditions in children of Neuquen, Patagonia, Argentina

Se investigó la prevalencia y distribución de parásitos intestinales (PI) en niños de 2 poblaciones de diferente nivel socioeconómico del área periurbana de la ciudad de Neuquén (Sectores I y II) a fin de evaluar su relación con las condiciones de hábitat y factores socio-económicos. Se procesaron muestras seriadas de materia fecal y de escobillado anal de 126 niños entre 2 y 14 años de edad. Se registraron datos acerca de condiciones de hábitat y factores socioeconómicos mediante visitas domiciliarias y encuestas observaciones estructuradas. Se detectó presencia de PI en el 50,7 por ciento de los niños del Sector I (barrio suburbano con adecuadas condiciones sanitarias y nivel socioeconómico medio o medio-bajo) y en el 92,9 por ciento de los niños del Sector II (asentamiento marginal con deficientes condiciones sanitarias y bajo nivel socioeconómico). Se identificaron 7 especies de protozoos intestinales y 4 especies de helmintos. Blastocystis hominis fue la especie más frecuente encontrada en ambas poblaciones. No se encontraron helmintos diferentes de Enterobius vermicularis en el Sector I y la prevalencia de tales especies fue muy baja en el Sector II. Las condiciones de hábitat deficientes y los bajos parámetros socioeconómicos se relacionaron con una mayor prevalencia de PI de transmisión directa como protozoos y E. vermicularis en las poblaciones estudiadas. Sin embargo, aún en ese contexto favorable a la transmisión, las especies parasitarias que requieren estadíos intermedios de maduración en el suelo no encuentran un hábitat adecuado para su diseminación en esta región patagónica.

The prevalence and distribution of intestinal parasites (IP) were investigated in children from two populations of different socioeconomic level, located in the same area of the city of Neuquén, in order to evaluate their relationship with habitat conditions and socioeconomic factors. Serial samples of faeces and anal scraping of 126 children between 2 and 14 years from two sectors of the suburban area of Neuquen (Sector I and Sector II) were analyzed. Data concerning habitat conditions and socioeconomic parameters were obtained by home visits and an observational structured survey. Presence of IP was detected in 50.7% of children from Sector I (suburban neighborhood with adequate sanitary conditions and middle or middle low socioeconomic level) and in 92.9% from children of Sector II (marginal settlement with poor sanitary conditions and low socioeconomic status). Seven intestinal protozoan and 4 helminth species were identified. Blastocystis hominis was the most frequent species found in both populations. No helminths different from Enterobius vermicularis were found in Sector I and the prevalence of such species was very low in Sector II. Deficient habitat conditions and low socioeconomic parameters showed relation with a higher prevalence of IP of direct transmission as protozoan and E.vermicularis in the studied populations. Nevertheless, even in this context favourable to transmission, the parasitic species which require intermediate stages of development in soil, don't find an adequate habitat for dissemination in this region