Clinical and laboratory features of rickettsioses in Yucatan, Mexico.

Rickettsia species are obligate intracellular bacteria that can cause mild to severe human disease. Based on phylogeny, clinical symptoms, and antigenic properties, rickettsiae are classified into four groups. Infections by these agents are characterized by clinical symptoms ranging from self-limited to severe and even fatal febrile illnesses, depending on the Rickettsia spp. involved, the patient's predisposition, and timely medical care. The present study aimed to characterize rickettsial diseases in Yucatan according to clinical and laboratory features appearing in medical records corresponding to 427 samples taken between 2015 and 2018. A study was conducted over the period 2015-2018 on 427 samples. Clinical and laboratory features were documented from the patients' medical records. For molecular diagnosis, blood was collected in 3.8 % sodium citrate as anticoagulant, and DNA was extracted. Single-step and nested PCR amplification was performed using genus-specific primers for the rickettsial 17kDa and ompB genes. The amplicons obtained were purified and sequenced. A total of 22.7 % (97/427) positive cases of Rickettsia spp. were identified by PCR from 14.15 % (15/106) of the municipalities in Yucatan. 75.2 % (73/97) of the cases were from the city of Merida during the autumn (September-December). The age groups with the highest frequency of confirmed cases were pediatric (5-14 years) (57.7 %) and adults (25-49 years) (42.2 %). There were six fatal cases in children, one associated with R. typhi and five with R. rickettsii. In non-fatal cases, 32.9 % (32/97) corresponded to the spotted fever group (SFG), and 60.8 % (59/97) to the typhus group (TG). Significant differences in signs, and laboratory data, were observed between the pediatric and adult populations. For the treatment of patients, oral and intravenous doxycycline was used in severe hospitalized cases. Typhus group and spotted fever group Rickettsiae are endemic pathological agents found in urban and rural areas of our region. Molecular identification allows for greater diagnostic accuracy and timely treatment and consequently a better prognosis. It is necessary to implement or reinforce measures focused on the dissemination of knowledge regarding rickettsial diseases and their prevention.

Design and evaluation of a mobile-based intervention for Maya adults during the COVID-19 pandemic

Introduction: The COVID-19 pandemic has had a direct impact on mental health. Inter national organisations have emphasised the vulnerability of indigenous people. Digital Mental Health approaches deliver online therapy as an evidence-based, effective, and accessible treat ment option for common mental health problems. However, the evidence regarding these ap proaches is limited in indigenous populations. The objective of this study is to describe the design, development, and evaluation of the efficacy of a self-applied online intervention regarding the psychological symptoms of depression, anxiety, and fear of COVID-19 in a sample of the Maya population. Method: A prospective longitudinal quantitative study was designed, where a single group was measured before and after receiving the online intervention. This study took place from April to September 2021 and consisted of six sessions delivered via WhatsApp in Spanish and Mayan. Results: The initial assessment was implemented with 82 participants who were evaluated using the Patient Health Questionnaire, Scale for Generalised Anxiety Disorder and the Fear of COVID-19 Scale; 18 participants remained in the intervention for the post-as sessment. Statistical differences were observed in PRE and POST measures of depression and anxiety, but not in fear of COVID-19. Conclusions: This study produced positive results for the first online mental health intervention implemented in the Latin American indigenous pop ulation. Future studies might consider developing similar interventions for other indigenous communities in Latin America.

Introducción: La pandemia de COVID-19 tuvo impacto directo en la salud mental. Organizaciones internacionales han enfatizado la vulnerabilidad de los pueblos indígenas. Los enfoques de salud mental digital brindan terapia en línea como una opción de tratamiento basada en evidencia, efectiva y accesible; sin embargo, los datos son limitados en población indígena. El objetivo de este estudio fue describir el diseño, desarrollo y evaluación de la eficacia de una intervención en línea autoaplicada sobre síntomas psicológicos de depresión, ansiedad y miedo al COVID-19 en una muestra de población maya. Método: Se diseñó un es tudio cuantitativo longitudinal prospectivo, donde se midió a un solo grupo antes y después de recibir la intervención en línea, implementada de abril a septiembre de 2021, que constó de seis sesiones impartidas vía WhatsApp, en español y maya. Resultados: La evaluación inicial se implementó con 82 participantes que fueron evaluados mediante el Cuestionario de Salud del Paciente, Escala para el Trastorno de Ansiedad Generalizada y Escala de Miedo al COVID-19; 18 participantes permanecieron para la evaluación posterior. Se observaron di ferencias estadísticas en las medidas pre- y post- de depresión y ansiedad, pero no miedo al COVID-19. Conclusiones: Este estudio arrojó resultados positivos de la primera intervención de salud mental en línea implementada en la población indígena latinoamericana. Estudios futuros podrían considerar el desarrollo de intervenciones similares para otras comunidades indígenas en América Latina.

Tick-associated diseases identified from hunting dogs during the COVID-19 pandemic in a Mayan community in Yucatan, Mexico.

Background: Hunting activity in the Mayan communities has increased due to COVID-19 and domestic dogs have gained more importance. Due to their proximity to humans, domestic dogs are a bridge between tick-borne diseases (TBDs) and humans and their peri-domestic environment. In Mexico, and especially in rural regions, there were not adequate records of TBDs during the SARS-CoV-2 pandemic. Aim: Identify TBD of ticks collected during the COVID-19 pandemic in a rural community. Methods: Tick capture was carried out in March 2021, in Teabo, Yucatan. Ticks were removed using from domestic dogs and placed in ethanol. Collected ticks were morphologically identified and underwent DNA extraction and a partial segment of the mitochondrial 16S-rDNA gene was amplified to corroborate the tick species. The DNA was screened for the presence of Anaplasma spp., Borrelia spp., Ehrlichia spp., and Rickettsia spp. Purified amplification products were submitted for sequencing and the results were compared to those deposited in GenBank using BLAST. Results: We collected 33 ectoparasites, Ixodes affinis, Rhipicephalus sanguineus, Rhipicephalus microplus, and Amblyomma mixtum on 11 hunting dogs. The most frequent ectoparasite was R. sanguineus (66%). We detected the presence of DNA of Rickettsia endosymbiont in I. affinis and Anaplasma platys in R. sanguineus. Rickettsia endosymbiont presented a similarity of 100% with the partial sequence of R. endosymbiont of I. affinis isolate IACACTM001 16S ribosomal RNA gene and the sequence of A. platys had a similarity of 100% with the partial sequence of the isolate 23-33TX 16S ribosomal RNA gene of A. platys from dogs from Texas, USA and with the partial sequence of the isolate L134 16S ribosomal RNA gene of Ehrlichia canis from dogs from Piura, Peru. Conclusion: We confirmed for the first time the presence of A. platys in R. sanguineus and R. endosymbiont in I. affinis ticks from dogs in the state of Yucatan.

Perception of risk among children: Exploring the risk of TB-rickettsial disease based on the children's drawing pictures in a Mayan community of Yucatan.

Background: It is important to work on designs of health promotion strategies that involve educational interventions about the risk factors associated to TB-rickettsiosis (Tick-Borne rickettsiosis). Children's drawings provide a window into their thoughts and feelings. The aim was to analyze children's risk perceptions regarding by the presence of ticks in a rural community in southeast Mexico. Methods: The main framework used was a social cognitive perspective under the Health Belief Model. Study was carried out in rural elementary schools and included a drawing contest. A total of 224 children (8-12 years old) participated. Drawings were coded and classified; descriptive trend analysis was performed using counts and percentages. The qualitative data was analyzed by researcher experts in TB- rickettsiosis and using Atlas ti V.8. Results: Results are presented in seven categories that consider environmental elements, actors, and cognitive aspects. The environmental context, 99.1% of the drawings represent sites outside the home (92% include vegetation with the presence of ticks in their immediate external environment). In the actors' category, 69.6% included people carrying out activities (prevention and risk). As for the cognitive aspects, 70% included domestic pets and 13.4% farmyards animals. Conclusion: The children expressed their ideas about risk factors and preventive measures against TB-rickettsiosis with a self-care approach. They acquired knowledge about preventive strategies and clinical symptoms. It is necessary to evaluate what happens after an intervention and how they implement in their lives what they have learned.

Rickettsia Vaccine Candidate pVAX1-OmpB24 Stimulates TCD4+INF-γ+ and TCD8+INF-γ+ Lymphocytes in Autologous Co-Culture of Human Cells.

BACKGROUND: In recent years, promising vaccination strategies against rickettsiosis have been described in experimental animal models and human cells. OmpB is considered an immunodominant antigen that is recognized by T and B cells. The aim of this study was to identify TCD4+INF-γ+ and TCD8+INF-γ+ lymphocytes in an autologous system with macrophages transfected with the vaccine candidate pVAX1-OmpB24. Lymphocytes and monocytes from 14 patients with Rickettsia were isolated from whole blood. Monocytes were differentiated into macrophages and transfected with the plasmid pVAX1-OmpB24 pVax1. Isolated lymphocytes were cultured with transfected macrophages. IFN-γ-producing TCD4+ and TCD8+ lymphocyte subpopulations were identified by flow cytometry, as was the percentage of macrophages expressing CD40+, CD80+, HLA-I and HLA-II. Also, we analyzed the exhausted condition of the T lymphocyte subpopulation by PD1 expression. Macrophages transfected with pVAX1-OmpB24 stimulated TCD4+INF-γ+ cells in healthy subjects and patients infected with R. typhi. Macrophages stimulated TCD8+INF-γ+ cells in healthy subjects and patients infected with R. rickettsii and R. felis. Cells from healthy donors stimulated with OmpB-24 showed a higher percentage of TCD4+PD1+. Cells from patients infected with R. rickettsii had a higher percentage of TCD8+PD-1+, and for those infected with R. typhi the larger number of cells corresponded to TCD4+PD1+. Human macrophages transfected with pVAX1-OmpB24 activated TCD4+IFN-γ+ and CD8+IFN-γ+ in patients infected with different Rickettsia species. However, PD1 expression played an important role in the inhibition of T lymphocytes with R. felis.

Fatal murine typhus with hemophagocytic lymphohistiocytosis in a child.

Hemophagocytic lymphohistiocytosis is a rare complication in Rickettsia typhi infections. We report the case of a 2-year-old boy with sudden night-onset fever, pallor, neck adenopathy and erythematous macular rash on the thorax, thighs and buttocks. During admission, he developed hyponatremia, hypoalbuminemia, anemia, thrombocytopenia, leukopenia, neutropenia, liver damage, hemorrhages and persistent fever. No hematological improvement was observed after the initial management, neoplastic diseases were discarded by bone marrow aspiration and lymph node biopsy; hemophagocytic lymphohistiocytosis was diagnosed. By immunohistochemistry and indirect immunofluorescence, murine typhus was also diagnosed and doxycycline was started with transitory recovery. Later, the child developed kidney failure and distributive shock that evolved to cardiac arrest and death. This is the first case report in Mexico on a fatal murine typhus associated with hemophagocytic lymphohistiocytosis in which the etiology was evidenced by histopathology.

Attitudes and Practices from People of a Mayan Community of Mexico, Related to Tick-Borne Diseases: Implications for the Design of Prevention Programs.

BACKGROUND: Tick-borne diseases are caused by several pathogens whose transmission could be associated to the life conditions of communities settled in endemic areas. We aimed to determine the knowledge, attitudes, and practices related to the exposition and prevention of tick-borne diseases among people living in a typical Mayan community of Yucatan, Mexico between Dec 2012 and May 2013. METHODS: A directed survey was applied to 212 (100%) householders (women and men) from Teabo, Yucatan, Mexico. Answers and field notes were recorded and analyzed with central statistics. RESULTS: People have been bitten at least once in the community, but the majority of them consider those bites innocuous. In addition, people do not consider prevention measures, and only a few mentioned the use of some chemicals on their backyards. CONCLUSION: This study found little awareness among the participants regarding the importance and the transmission of these diseases even though they possess a vast knowledge regarding ticks. Therefore, educational strategies and prevention programs that include these habits for its modification are required to minimize the exposition to the vectors.

Presence of Rickettsia Species in a Marginalized Area of Yucatan, Mexico.

In the state of Yucatan, Mexico, rickettsiosis has become a common vector-borne disease in the general population. Ectoparasite species such as Rhipicephalus sanguineus and Amblyomma mixtum have been identified as Rickettsia vectors in Yucatan by studies focused on the wild animal population in rural areas. There have been studies that have tried to determine the presence of Rickettsia species in ectoparasites collected in Yucatan, but these studies did not include marginalized areas, where living in close contact with domestic and peridomestic animals that carry ectoparasites is a high-risk factor for acquiring rickettsial infection or many other vector-borne diseases. We evaluated the vector diversity and the presence of Rickettsia species presence in the ectoparasite population that parasitizes domestic animals in a marginalized rural town of Yucatan, Mexico; we also evaluated the seroprevalence of rickettsial antibodies in the human population of this town in order to determine the prevalence of rickettsial infection. A total of 437 ectoparasites were collected from the study area. The tick specimens collected belonged to the species Rhipicephalus sanguineus (n=380, 49 positive), Amblyomma mixtum (n=3, 0 positive), Ixodes affinis (n=4, 0 positive), Ctenocephalides felis (n=33, 0 positive), and Trichodectes canis (n=17, 0 positive). Conventional polymerase chain reaction and sequencing were used to identify the DNA of Rickettsia. Six out of 354 (1.8%) serum samples were positive for antibody to R. typhi. The combination of low antibody titers and the presence of Rickettsia species infecting ectoparasite species found in the study area requires eco-epidemiological studies and the identification of potentially protective practices or habits.

Immunogenicity of OmpA and OmpB antigens from Rickettsia rickettsii on mononuclear cells from Rickettsia positive Mexican patients.

Background & objectives: The nature of the rickettsial antigens and the immune response generated by them, have been the subject of exhaustive research so that a suitable vaccine can be developed. Till date evaluations of Rickettsia rickettsii antigens that induce both humoral and cellular responses in animal models have only shown partial protection and short-term immunological memory. This study was aimed to evaluate the immune response induced by DNA plasmids generated from the OmpA and OmpB genes of R. rickettsii in peripheral blood mononuclear cells of rickettsial (sensitized) patients compared to healthy subjects. Methods: Plasmids OmpA-49, OmpB-15 and OmpB-24 were generated in the pVAX vector. Macrophages derived from the THP-1 cell line were transfected in vitro with the plasmids and were co-cultured with T-lymphocytes from sensitized subjects and healthy subjects to evaluate cell proliferation and cytokine production. Results: The OmpB-24 plasmid induced proliferative response in human lymphocytes, with production of IL-2, IFN-γ, IL-12p70, IL-6 and TNF-α, likely due to the presence of conserved epitopes among R. rickettsii, R. typhi and R. felis (differing from 1 to 3 amino acids) during the construction of the plasmids. Interpretation & conclusion: DNA sequences of rickettsial epitopes can be cloned into the pVAX vector. Constructed plasmids can generate a proliferative response and produce cytokines in vitro, in co-culture of transfected macrophages with sensitized human lymphocytes. Plasmid OmpB-24 proved to be the most immunogenic with respect to plasmids OmpA-49 and OmpB-15.

Mecanismos de resistencia antifúngica de los azoles en Candida albicans. Una revisión / Mechanisms of antifungal resistance of azoles in Candida albicans. A review

Resumen Candida albicans es una levadura comensal capaz de causar una infección oportunista en hospederos susceptibles denominada candidiasis. El tratamiento para combatir la candidiasis puede ser tópico o sistémico según el tipo de infección, los antifúngicos más utilizados son los derivados imidazólicos (fluconazol, itraconazol, ketoconazol, miconazol etc.), sin embargo en la actualidad se observa una disminución en la efectividad de estos medicamentos, es decir, un fenómeno de resistencia de parte del microorganismo a estos fármacos, esto debido principalmente, al surgimiento de levaduras resistentes, a la aparición de nuevas especies patógenas, a la prescripción irracional de antimicóticos como profilaxis y al aumento de las dosis terapéuticas. Existen dos mecanismos por los que Candida puede adquirir resistencia a un azol. El primero es por mutaciones moleculares de la enzima diana del antifúngico, como la alteración de las enzimas relacionadas en la síntesis del ergosterol y el segundo por la alteración en las bombas de expulsión: ATP-binding cassette (ABC) y facilitadores mayores (MF). En este trabajo se resumen los principales mecanismos de resistencia en Candida y la importancia de hacer pruebas de susceptibilidad con el fin de brindar un tratamiento adecuado para este tipo de infecciones oportunistas.

Abstract Candida albicans is a commensal yeast capable of causing an opportunistic infection called candidiasis in susceptible hosts. Treatment to combat Candida may be topical or systemic according to the type of infection and the imidazole derivatives (fluconazole, itraconazole, ketoconazole, miconazole, etc.) are the antifungals most widely used. However, resistance to these drugs is observed by a decrement in their effectiveness. This is mainly due to the emergence of resistant yeasts and of new pathogenic species, as well as to the irrational prescribing of antifungal prophylaxis and the use of higher therapeutic doses. There are two mechanisms by which Candida can acquire an azole resistance, the first is by molecular mutations of antifungal target enzyme, as the alteration of enzymes related to the synthesis of ergosterols and the second by change in the efflux pumps, such as those of ATP-binding cassette and the higher facilitators. In this work the main mechanisms of resistance to Candida and the importance of performing susceptibility tests in order to provide an adequate treatment for this type of opportunistic infections are summarized.

Rickettsia felis as emergent global threat for humans.

Rickettsia felis is an emergent pathogen belonging to transitional group rickettsiae. First described in 1990, R. felis infections have been reported to occur worldwide in fleas, mammals, and humans. Because clinical signs of the illness are similar to those of murine typhus and other febrile illnesses such as dengue, the infection in humans is likely underestimated. R. felis has been found throughout the world in several types of ectoparasites; cat fleas appear to be the most common vectors. R. felis infection should be considered an emergent threat to human health.