Prevalence and Progression of Gingivitis in Domestic Cats in Subtropical Mexico.

With the objective to characterize the gingival index (GI) and its progression, 218 domestic cats in a subtropical region of Mexico were studied. All teeth of each cat were examined with a periodontal probe to determine the GI; in addition, the absence of teeth was recorded. Six months later, the teeth of the 38 cats were again examined to assess any progression of the GI and loss of teeth. From the 218 cats, 33.0% of them develop some degree of gingival inflammation; from those, 61.5% were classified as GI 1. Age, sex, and neutered status were associated with tooth affections. Missed teeth were observed in 35% of the cats, particularly for molars 109 and 209 in both sexes. After six months, the number of teeth with GI 1 decreased to 20%. The gingival problems in cats have not been well studied, particularly at the speed they progress and how this can affect the loss of teeth; under the conditions of this study, a high frequency of gingival inflammation even at early age was demonstrated, with a rapid tooth loss. Although young males were more prone to develop gingivitis, females tend to loss more teeth. Non-neutered cats tended to develop more dental affections.

Congenital Transmission of Trypanosoma cruzi in Naturally Infected Dogs.

Background: Congenital transmission (CT) of Trypanosoma cruzi in dogs has not been clearly demonstrated, even though dogs are important reservoirs of this agent. Materials and Methods: Seventeen late pregnant dogs seropositive for T. cruzi were selected, and a total of 84 fetuses were obtained. Blood and heart tissues from the fetuses and dams, and placental tissue from dam were collected. All tissues were analyzed by quantitative polymerase chain reaction (qPCR) for T. cruzi DNA (TcDNA) and inflammatory infiltrate and pathology by histological examination. CT was determined when physical, histological, or molecular evidence of T. cruzi was detected in blood or tissues of the fetuses. Results: A general transmission frequency of 59% was found, and 0.20 ± 0.24 of fetuses per litter were infected. Dams that were qPCR positive for TcDNA in cardiac tissue or blood displayed a transmission frequency of 100% and 67%, respectively. The highest parasite burden was noted in dams that were positive for TcDNA in both blood (82E-01 ± 1.54E-01) and cardiac (5.28E+03 ± 8.85E+03) tissues. In fetuses, higher parasitic burden in blood and cardiac tissue was found in those carried by dams that were seropositive and qPCR positive for TcDNA in cardiac tissue and blood. No amastigote nests were recorded in the cardiac tissue of fetuses in the histopathological studies, but typical lesions of T. cruzi infection were identified in all fetuses where CT occurred. Conclusions: CT of T. cruzi occurred at a high frequency in naturally infected pregnant dogs from the endemic areas.

Impacto indirecto de COVID-19 en las tasas de incidencia de enfermedades transmitidas por vector en México / Indirect impact of COVID-19 on the incidence rates of vector-borne diseases in Mexico

Resumen Introducción: La distribución espacial y temporal de la infección por SARS-CoV-2 sobrepasa las áreas endémicas de enfermedades transmitidas por vector (ETV), cuya vigilancia en México ha cambiado sustancialmente a partir del primer caso confirmado de COVID-19. Objetivos: Estimar y comparar las tasas de incidencia de las ETV antes y después de la introducción del SARS-CoV-2 en México. Métodos: Estudio retrospectivo de casos de ETV de 2014 a 2021. Las tasas de incidencia de cada ETV en el periodo previo (2014-2019) y posterior (2020-2021) a la introducción del SARS-CoV-2 en México fueron calculadas y comparadas. Resultados: Antes de la introducción del SARS-CoV-2, las tasas de incidencia de las ETV fueron altas y posterior a la introducción del coronavirus hubo un descenso en los índices epidemiológicos; sin embargo, solo se identificó diferencia estadística significativa en la tasa de incidencia de la malaria (p ≤ 0.05) y otras rickettsias (p ≤ 0.05). Conclusiones: Algunas medidas para reducir los casos de COVID-19, como el distanciamiento social, el confinamiento domiciliario, la reducción en el aforo en el transporte público y el trabajo en casa, probablemente contribuyeron a disminuir temporalmente el número de casos de las ETV; sin embargo, puede haber rebrote de las ETV en el futuro cercano.

Abstract Introduction: SARS-CoV-2 infection spatial and temporal distribution overlaps with endemic areas of vector-borne diseases (VBD), whose surveillance in Mexico has substantially changed since the first COVID-19 confirmed case. Objectives: To estimate and compare the incidence rates of VBDs before and after the introduction of SARS-CoV-2 in Mexico. Methods: Retrospective study of VBD cases from 2014 to 2021. The incidence rates of each VBD in the period before (2014-2019) and after (2020-2021) the introduction of SARS-CoV-2 in Mexico were calculated and compared. Results: Before the introduction of SARS-CoV-2, the incidence rates of VBDs were high and after the introduction of coronavirus there was a decrease in epidemiological indices; however, there was only statistically significant difference in the incidence rate of malaria (p ≤ 0.05) and other rickettsiae (p ≤ 0.05). Conclusions: Some measures to reduce COVID-19 cases, such as social distancing, home confinement, reductions in public transport and working at home (home office), probably temporarily decreased the number of VBD cases; however, there may be a resurgence of VBDs in the near future.

Animal Models of Trypanosoma cruzi Congenital Transmission.

Chagas disease, initiated by the etiological agent Trypanosoma cruzi, is an endemic infection in the American continent. Although vectorial transmission of T. cruzi is recognized as the main mode of infection, other routes such as congenital and blood transfusion are also documented as important methods of transmission. T. cruzi maternal-fetal transmission has been recorded in humans and examined by some investigators in naturally and experimentally infected mammals. Dogs are recognized as the major reservoir host in maintaining the domestic transmission of T. cruzi; however, the importance of congenital transmission in preserving the infection cycle in dogs has not been studied in detail. In this article, we reviewed the current knowledge of congenital transmission of T. cruzi in humans and compared the placental architecture of humans and different animals with particular attention to rodents, dogs, and non-human primates that have been used as experimental models of T. cruzi infection, congenital transmission, and Chagas disease pathogenesis. The placentas of humans and animals have some similar and dissimilar characteristics that should inform the study design and interpretation of results when evaluating the efficacy of new anti-parasite drugs and therapies against congenital infection.

Quantification of Osteoprotegerin Plasma Levels in Patients with Periodontitis.

OBJECTIVE: Periodontitis (POD) is an infectious process directed at the structures supporting the teeth. Destruction of alveolar bone is considered one of the main causes of tooth loss in humans and is mediated by the host immune response. Osteoprotegerin (OPG), a protein that inhibits bone resorption by binding to the RANK ligand (RANKL), prevents osteoclastic differentiation. The aim of the study was to determine the plasma levels of OPG in patients with POD. METHODS: a case-control study with forty-nine patients with POD and 49 healthy controls were included in the study. OPG levels were determined by an ELISA test in plasma samples. RESULTS: OPG values (1.6203 ng/mL) were higher in the POD group compared with control group (1.2824 ng/mL). Among the studied groups, we detected significant differences in age, glycosylated haemoglobin (HbA1C), and plasma concentration of OPG (p < 0.05). CONCLUSION: plasma OPG levels are associated with bone formation and destruction processes, suggesting that OPG acts in a protective manner.

Quantification of sphingosine-1-phosphate (S1P) in patients with periodontitis stage II grade B / Cuantificación de esfingosina 1 fosfato en pacientes con periodontitis estadío II grado B

Abstract The aim of this study was the quantification of Sphingosine-1-phosphate (S1P) in periodontal pockets of patients with periodontitis. This is an observational, descriptive, case-control study. Thirty subjects were selected: 15 controls and 15 cases. A periodontal study was conducted following the parameters of AAP 2017 for the diagnosis of periodontal diseases. A sample of saliva and gingival crevicular fluid was obtained from each subject and then analyzed with the Human S1P Elisa kit (MyBioSource #MBS2516132) accordingly to the manufacturer's instructions, in order to verify the presence of S1P and quantify it´s concentration when founded. Results showed a significant difference (p=0.05) between cases and controls. In the case of saliva samples, the concentration of S1P was higher than the ones found in the control group (72.94 ng/mL and 45.12 ng/mL). For GCF, a higher amount of S1P was found in patients with POD (20.09 ng/mL and 15.20 ng/mL). This work raises a possible route of bone metabolism, inflammatory process, and identification of periodontitis through oral quantification of S1P, however, future studies are needed.

Resumen El propósito de este estudio fue la cuantificación de Esfingosina-1-Fosfato (S1P) en las bolsas periodontales de pacientes con periodontitis. Estudio observacional, descriptivo de casos y controles. 30 sujetos fueron seleccionados de los cuales 15 controles y 15 casos. Se realizó un estudio periodontal completo siguiendo los parámetros establecidos por la AAP en 2017 para el diagnóstico de las enfermedades periodontales. Se tomaron muestras de saliva y de líquido crevicular gingival de cada sujeto estudiado y se analizaron con el ELISA kit humano para S1P (MyBiosource #MBS2516132) y de acuerdo con las instrucciones del fabricante, se realizó para cuantificar la presencia d S1P en las muestras estudiadas. Los resultados mostraron diferencia significativa (p=0.05) entre casos y controles. En el caso de las muestras de saliva, la concentración de S1P en controles fue mayor (72.94 ng/mL y 45.12 ng/mL). Para Líquido crevicular gingival, se encontró mayor cantidad de S1P en los pacientes con periodontitis (20.09 ng/mL y 15.20 ng/mL). Este estudio plantea una posible ruta de metabolismo óseo, proceso inflamatorio e identificación de la Periodontitis a través de la cuantificación oral de S1P, sin embargo se necesitan estudios futuros.

Combined use of real-time PCR and serological techniques for improved surveillance of chronic and acute American trypanosomiasis in dogs and their owners from an endemic rural area of Neotropical Mexico.

In this study, the prevalence of T. cruzi infection was estimated in dogs and their owners from a rural community in Mexico using serological techniques for chronic infection cases, qPCR for acute phase cases, and a combination of both techniques to detect chronic and acute infections. Eighty-nine blood samples were collected from owners and their dogs for obtaining serum and parasite DNA. Prevalence was calculated using (i) positive cases detected in a serological test (ELISA and Western blot), (ii) positive cases detected in a qPCR test, and (iii) positive cases detected by both techniques. Sensitivity, specificity, and predictive values were determined individually for serology, qPCR and for both techniques used simultaneously. The prevalence observed varied: for serology, 25.8% of the dogs and 7.9% of the owners were seropositive, while for qPCR 29.2% of the dogs and 10.1% of the owners were identified as positive. Combination of serological and molecular techniques resulted in a prevalence of 38.2% for dogs and 12.4% for their owners. The sensitivity, specificity and predictive values calculated for both techniques improved when both techniques were used simultaneously (sensitivity of 92.4% and specificity of 100% for infected dogs and sensitivity of 93.4% and specificity of 100% for infected owners). Combined use of serological tests and qPCR allowed identifying a greater number of positive cases in dogs and their owners. This strategy may help implement adequate and timely epidemiological surveillance of American trypanosomiasis in order to prevent the appearance of new cases of Trypanosoma cruzi infections in endemic zones.

Evaluation of the effectiveness of fluralaner against adult stages of Rhodnius prolixus in dogs.

Triatomines are vectors of American Trypanosomiasis also known as Chagas´ disease where several reservoirs including dogs are involved in the transmission cycle of the causal agent (Trypanosoma cruzi). Considering that the prevalence of American trypanosomiasis in dogs is higher than in humans and that dogs in addition are susceptible of this disease, and are involved in peridomestic transmission to humans, the search for new alternatives for vector control of the triatomines responsible for transmission in dogs is required. Over the 20 weeks the study lasted, 600 individual female, adult of Rhodnius prolixus were offered to the 15 dogs treated with a single oral dose of Fluralaner (Bravecto®, MSD). Feeding pattern of triatomines was not affected by the treatment during the whole study. The fluralaner-induced mortality of R. prolixus had a significant effect until week 12 at which time 100% mortality was observed. Mortality decreased to 67.5% at week 16 to practically nil 0.8% on week 20. Fluralaner achieved 100% mortality of triatomines between 12- and 48-h post-feeding. It was demonstrated that a single oral dose of fluralaner in dogs is highly effective in producing mortality in adult R. prolixus for the time guaranteed by the manufacturer for other blood-sucking insects, with a considerable effective residual effect for up to 16 weeks. Due to this high efficacy, fluralaner could be considered in strategies to control the transmission vectors of Chagas disease in dogs and in turn decrease the peri-domestic transmission cycle, particularly in hyperendemic areas.

Indirect impact of COVID-19 on the incidence rates of vector-borne diseases in Mexico.

INTRODUCTION: SARS-CoV-2 infection spatial and temporal distribution overlaps with endemic areas of vector-borne diseases (VBD), whose surveillance in Mexico has substantially changed since the first COVID-19 confirmed case. OBJECTIVES: To estimate and compare the incidence rates of VBDs before and after the introduction of SARS-CoV-2 in Mexico. METHODS: Retrospective study of VBD cases from 2014 to 2021. The incidence rates of each VBD in the period before (2014-2019) and after (2020-2021) the introduction of SARS-CoV-2 in Mexico were calculated and compared. RESULTS: Before the introduction of SARS-CoV-2, the incidence rates of VBDs were high and after the introduction of coronavirus there was a decrease in epidemiological indices; however, there was only statistically significant difference in the incidence rate of malaria (p ≤ 0.05) and other rickettsiae (p ≤ 0.05). CONCLUSIONS: Some measures to reduce COVID-19 cases, such as social distancing, home confinement, reductions in public transport and working at home (home office), probably temporarily decreased the number of VBD cases; however, there may be a resurgence of VBDs in the near future.

INTRODUCCIÓN: La distribución espacial y temporal de la infección por SARS-CoV-2 sobrepasa las áreas endémicas de enfermedades transmitidas por vector (ETV), cuya vigilancia en México ha cambiado sustancialmente a partir del primer caso confirmado de COVID-19. OBJETIVOS: Estimar y comparar las tasas de incidencia de las ETV antes y después de la introducción del SARS-CoV-2 en México. MÉTODOS: Estudio retrospectivo de casos de ETV de 2014 a 2021. Las tasas de incidencia de cada ETV en el periodo previo (2014-2019) y posterior (2020-2021) a la introducción del SARS-CoV-2 en México fueron calculadas y comparadas. RESULTADOS: Antes de la introducción del SARS-CoV-2, las tasas de incidencia de las ETV fueron altas y posterior a la introducción del coronavirus hubo un descenso en los índices epidemiológicos; sin embargo, solo se identificó diferencia estadística significativa en la tasa de incidencia de la malaria (p ≤ 0.05) y otras rickettsias (p ≤ 0.05). CONCLUSIONES: Algunas medidas para reducir los casos de COVID-19, como el distanciamiento social, el confinamiento domiciliario, la reducción en el aforo en el transporte público y el trabajo en casa, probablemente contribuyeron a disminuir temporalmente el número de casos de las ETV; sin embargo, puede haber rebrote de las ETV en el futuro cercano.

Four Species of Under-reported Parasitic Arthropods in Mexico and their Potential Role as Vectors of Pathogens.

Numerous arthropod taxa are important in human and veterinary medicine. The salivary secretions and feces of arthropods can cause allergic reactions in host vertebrates or harbor pathogens. Also, bites can be a risk factor for secondary infections. Documenting the diversity of arthropods of medical and veterinary importance remains an important aspect of disease control and prevention. We provide new records of ectoparasitic arthropods from Mexico that are of potential medical or veterinary relevance. Scanning electron microscopy along with amplification and sequencing of a fragment of the mitochondrial gene (16S rRNA) was used to confirm some species identities. We report the cat louse Felicola subrostratus from cats and the chewing louse Heterodoxus spiniger from dogs, which are common ectoparasites but largely not reported in Mexico. The chigger Eutrombicula alfreddugesi is common on wild lizards (Squamata). For the first time, E. alfreddugesi is reported on Hemidactylus frenatus (common house gecko). This reptile has a close relationship with humans and its chiggers can cause dermatitis (i.e., trombiculiasis) or transmit pathogens. In addition, the common bed bug Cimex lectularius is reported for the first time in the state of Yucatan, an atypical area for its natural distribution. Phylogenetic analysis revealed that Cimex lectularius from Yucatan is closely related to genetic sequences of Cimex lectularius from China. Knowing the regional distribution of arthropods allows the design and implementation of prevention strategies for those that have potential roles as reservoirs or vectors.

Presence of congenital anomalies in three dog litters.

This report describes multiple congenital malformations found in three dog litters delivered by emergency caesarean section. In all of the litters, some puppies were born alive but were euthanized because of the seriousness of their malformations and low probability of survival. In two litters, gastroschisis was associated with amelia of the right anterior limb. Other malformations such as anencephaly were also found in three puppies among the different litters. This report describes the morphological findings of the affected puppies, discusses the most appropriate terminologies for each case and highlights the importance of an epidemiological survey to identify potential factors associated with the cases.

A fatal case of canine cutaneous leishmaniosis in a dog

A 10-year-old intact female Chihuahua, 2.5 kg of weight and BCS 2 (range 1­5) was taken in for medical consultation due to the presence of four skin lesion, two in the ventral thoracic region and two in the dorsal region. The dog was receiving medication due to congestive heart failure. A cutaneous form of canine leishmaniosis was diagnosed using molecular tools from a 10-years-old Chihuahua dog living in the Caribbean region. The critical health condition of the geriatric patient may have evolved to a fatal renal failure. This report is the first of a fatal case of leishmaniosis in a dog from the endemic region in Mexico.

Screening of Zoonotic Parasites in Playground Sandboxes of Public Parks from Subtropical Mexico.

The pathological agents Toxoplasma gondii, Ancylostoma caninum, and Toxocara canis are widely distributed zoonotic parasites with high prevalence in tropical and subtropical regions of the world. The aim of the present study was to determine the presence of DNA from these parasites in sand samples from the sand playgrounds in the southeastern region of Mexico. Samples of sand were collected from 68 playgrounds in public parks in the city of Merida, Yucatan, which is the main urban area in the southeast of Mexico. The samples were examined using nested PCR to detect the SAG1 gene from Toxoplasma gondii, and endpoint PCR for the amplification of ITS-2 and rRNA-ITS2 genes from Toxocara canis and Ancylostoma caninum, respectively. The presence of T. gondii DNA was detected in 11.8% (8/68) samples, DNA from A. caninum and T. canis was not detected. Results indicate that playgrounds from the studied sandboxes are contaminated with T. gondii oocysts and may represent a risk of infection for people in contact with the sand, especially for preschoolers.

Quantification of TNF-α in Patients with Periodontitis and Type 2 Diabetes.

OBJECTIVE: The present study aimed to compare variations in quantified tumor necrosis factor-alpha (TNF-α) levels in patients with periodontitis stage 2 grade B (POD2B) and/or type 2 diabetes (T2D) and to identify any relationships between this cytokine and these diseases. METHODS: Levels of the cytokine TNF-α in gingival crevicular fluid in patients with POD2B and/or T2D were evaluated. A total of 160 subjects were distributed into four groups: those with POD2B (n=44); those with T2D (n=37); those with POD2B/T2D (n=40); and healthy subjects (n=39). Glycosylated hemoglobin (HbA1c) and blood glucose (BG) levels were quantified in each subject. Data were collected on body mass index (BMI), loss of insertion (LI), and probe depth (PD). Gingival crevicular fluid samples were collected from the most acutely affected periodontal pocket and gingival sulcus in each subject, and TNF-α was quantified by multiplex analysis. RESULTS: Kruskal Wallis tests was used to identify differences in TNF-α levels, LI, PD, BMI, BG, and HbA1c by group. Differences (p<0.001) were found for LI, PD, BG, and HbA1c. A Spearman test was used to calculate possible correlations between TNF-α levels and LI or PD identified a weak but significant negative correlation of TNF-α with LI (Rho=-0199; p=0.012), and a moderately positive correlation of LI with PD (Rho=0.509; p < 0.001). CONCLUSIONS: No variation was found between TNF-α levels and the presence of POD2B, POD2B/T2D, or T2D, suggesting the absence of any direct relationship between progression of these diseases and TNF-α levels. However, a correlation was present between low TNF-α concentrations and greater LI.

Toxoplasma gondii in Captive Wild Felids of Mexico: Its Frequency and Capability to Eliminate Oocysts.

There is little information about Toxoplasma gondii in wild felids, even when these species have been associated with cases of toxoplasmosis in humans. In this study, samples of serum and whole blood were collected from 42 felids from 10 different species, in 4 Mexican zoos. Stool samples from 36 animals were also collected, corresponding to 82% of the felids included in the study. Stool samples were used for the search of oocysts by light field microscopy and PCR. Serum samples were analyzed by indirect immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) and indirect fluorescent antibody test (IFAT). DNA samples were purified from whole blood and stool for the amplification of a fragment of the SAG1 gene of T. gondii by a nested PCR (nPCR). The seroprevalence of IgG anti-T. gondii-specific antibodies by means of the ELISA was 100% (42/42) and 52.4% (22/42) by IFAT. The titers obtained varied from 1:80 to 1:2560. DNA of T. gondii was detected in 9.5% (4/42) of the blood samples by using nPCR. No oocysts were observed in the stool samples analyzed by light field microscopy. However, the DNA of the parasite was identified in 14.3% (5/35) of the stool samples evaluated. These results indicate a high prevalence of T. gondii in the studied populations of wild felids in captivity, with evidence of parasitemia and elimination of few oocysts even in adult hosts.

Frequency of Trypanosoma cruzi Infection in Synanthropic and Wild Rodents Captured in a Rural Community in Southeast of Mexico.

The protozoan parasite Trypanosoma cruzi is the causative agent of the Chagas disease, which is endemic in southeastern Mexico and is transmitted by the vector Triatoma dimidiata (triatomide). T. cruzi infect a great variety of domestic and wild mammals; rodents are considered one of the most important reservoirs of the parasite in the transmission cycles of T. cruzi. The objective of this study was to determine the frequency of T. cruzi infection and to determine the parasitic load in synanthropic and wild rodents from the rural community of southern Mexico. A total of 41 blood samples and 68 heart tissue samples were collected from various species of synanthropic (n= 48 in 2 species) and wild rodents (n= 35 in 5 species). DNA was extracted from samples to detect the presence of T. cruzi through quantitative PCR (qPCR). T. cruzi DNA was detected in the 9.75% of the blood samples of the synanthropic species (4/41) (14.28%) for Rattus rattus samples and 25% for Ototylomys phyllotis samples, with an average of parasitic load of 4.80 ± 1.17 parasites/µL. In the case of heart tissue samples, 10.29% were positive for T. cruzi (7/68) (8.7% for Rattus rattus, 40% for Peromyscus yucatanicus, and 42.8% for Ototylomys phyllotis) with an average parasite load of 3.15 ± 1.98 eq-parasites/mg. The active and chronic infection of T. cruzi in synanthropic or wild rodents of the rural community of southern Mexico evidences the natural infection in these reservoirs which contribute to maintaining the agent in the wild and domestic environments and can represent a risk of infection for the human population when the vector is present.

Stray Dog Population in a City of Southern Mexico and Its Impact on the Contamination of Public Areas.

To assess the risk of zoonotic pathogen transmission as function of stray dog presence and health status, a cross-sectional study was carried out in a large city of southern Mexico that lacks comprehensive strategies for the control of stray canine populations. The photographic capture-recapture method was used to estimate the density of dogs/km2. In the same way, dog feces from 14 public parks of the city were collected to determine the prevalence and intensity of infection with gastrointestinal parasites. The canine population was estimated between 65 and 80 thousand dogs, with a population density of 1,081 dogs/km2, mostly males (71.4%). A high proportion of dogs (72.3%) were found to be in good body condition score (BCS 3). The person:dog ratio was 2.3. The likelihood of being in the BCS 2 category was lower in areas with a higher density of dogs. All feces collected from the parks contained eggs of intestinal parasites, most of them with a medium (42.9%) to high (35.7%) infection intensity, notably Ancylostoma caninum. It was recorded that cases with a low-intensity of GI infection showed polyparasitism (35.7%) associated with A. caninum. There is a large population of stray dogs that roam freely in the streets of Campeche city with access to sources of food, which is reflected by their good BCS, and dogs do not have access to preventive medicine programs (de-worming) and thus contaminate public parks with feces with significant parasitic egg loads of zoonotic importance.

Efficacy of Recombinase Polymerase Amplification to Diagnose Trypanosoma cruzi Infection in Dogs with Cardiac Alterations from an Endemic Area of Mexico.

Chagas disease is a lingering Public Health problem in Latin America with ∼5.7 million people infected with Trypanosoma cruzi. Transmission is still taking place in most countries of the Americas, including the United States. Dogs are frequently infected with T. cruzi and its high infection prevalence is associated with increased risk of Chagas disease in humans. The city of Mérida in the Yucatan peninsula is endemic for Chagas disease and canines are frequently infected with T. cruzi. The objective of this study was to evaluate the performance of a qualitative point of care (POC) molecular test (RPA-LF, recombinase polymerase amplification-lateral flow) developed in our laboratory for identifying infected dogs. We used retrospective samples of dogs that came for consultation because of cardiac alterations and proved to be infected with T. cruzi as determined by enzyme-linked immunosorbent assay (ELISA), Western blot, and quantitative PCR (qPCR). The analytical sensitivity indicated that RPA-LF amplified T. cruzi DNA in samples containing almost equal to one to two parasites per reaction. Serial twofold dilutions of T. cruzi epimastigotes showed that the test had 95% (19/20) repeatability at concentrations of two parasites per reaction. The test showed no cross reactivity with human DNA or other protozoan parasites (Trypanosoma rangeli, Leishmania spp., and Plasmodium spp.). RPA-LF had the capacity to amplify all discrete typing units (DTUs I-VI) of T. cruzi that circulate in domestic or extradomestic environments. The RPA-LF had 93.2% (95% confidence interval 87.2-98.1) sensitivity and excellent agreement with qPCR used as gold standard (Cohen's Kappa test = 0.963). ELISA was positive in 96.6% (85/88) of dogs, which together with the molecular tests confirmed the frequent contact with infected triatomine bugs in the city of Mérida. These preliminary results on the diagnostic efficacy of the RPA-LF deserve further large-scale field testing of this POC test for T. cruzi infection in endemic areas.

Quantitative and histological assessment of maternal-fetal transmission of Trypanosoma cruzi in guinea pigs: An experimental model of congenital Chagas disease.

OBJECTIVE: We evaluated the effect of Trypanosoma cruzi infection on fertility, gestation outcome, and maternal-fetal transmission in guinea pigs (Cavia porcellus). METHODS: Animals were infected with T. cruzi H4 strain (TcI lineage) before gestation (IBG) or during gestation (IDG). Tissue and sera samples of dams and fetuses were obtained near parturition. RESULTS: All IBG and IDG dams were seropositive by two tests, and exhibited blood parasite load of 1.62±2.2 and 50.1±62 parasites/µl, respectively, by quantitative PCR. Histological evaluation showed muscle fiber degeneration and cellular necrosis in all infected dams. Parasite nests were not detected in infected dams by histology. However, qPCR analysis detected parasites-eq/g heart tissue of 153±104.7 and 169.3±129.4 in IBG and IDG dams, respectively. All fetuses of infected dams were positive for anti-parasite IgG antibodies and tissue parasites by qPCR, but presented a low level of tissue inflammatory infiltrate. Fetuses of IDG (vs. IBG) dams exhibited higher degree of muscle fiber degeneration and cellular necrosis in the heart and skeletal tissues. The placental tissue exhibited no inflammatory lesions and amastigote nests, yet parasites-eq/g of 381.2±34.3 and 79.2±84.9 were detected in IDG and IBG placentas, respectively. Fetal development was compromised, and evidenced by a decline in weight, crow-rump length, and abdominal width in both groups. CONCLUSIONS: T. cruzi TcI has a high capacity of congenital transmission even when it was inoculated at a very low dose before or during gestation. Tissue lesions, parasite load, and fetal under development provide evidence for high virulence of the parasite during pregnancy. Despite finding of high parasite burden by qPCR, placentas were protected from cellular damage. Our studies offer an experimental model to study the efficacy of vaccines and drugs against congenital transmission of T. cruzi. These results also call for T. cruzi screening in pregnant women and adequate follow up of the newborns in endemic areas.