Influenza virus and proteolytic bacteria co-infection in respiratory tract from individuals presenting respiratory manifestations

A role for proteolytic bacteria in the exacerbation of influenza virus has been shown in natural hosts such as pigs and humans. Four hundred seven samples were collected from the respiratory tract of individuals presenting clinical manifestations, during influenza season (2003-2005) in São Paulo City. The aim of this study was to evaluate the incidence of determined bacteria co-infecting virus in human respiratory tract. Tests, such as bacteriological, immunofluorescence (IF), RT/PCR and hemagglutination (HA) were used for bacterial and viral investigation. Thirty seven (9.09%) positive for influenza virus were screened by IF. The RT/PCR confirmed the presence of influenza virus in these samples. Bacterial and agar casein tests demonstrated that 18 (48.64%) individuals were infected with proteolytic bacteria such as Staphylococcus spp., Streptococcus spp. and Pseudomonas spp. Among these samples, 13 (35.13%) were co-infected with influenza A virus. Influenza type B, co-infecting bacteria were found in five (13.51%) samples. In vitro the S. aureus protease increased the influenza HA titer after contact for 30 min at 25 ºC. Results revealed the occurrence of co-infection with proteolytic bacteria and influenza in the evaluated individuals. This finding corroborates that virus versus bacteria synergism could be able to potentiate respiratory infection, increasing damage to hosts.

Preliminary investigation of Culicidae species in South Pantanal, Brazil and their potential importance in arbovirus transmission

In view of the high circulation of migratory birds and the environmental and climatic conditions which favor the proliferation of arthropods, the Brazilian Pantanal is susceptible to circulation of arboviruses. However, the amount of data concerning arbovirus vectors in this area is scarce; therefore the aim of this study was to conduct a preliminary investigation of Culicidae species in the Nhecolândia Sub-region of South Pantanal, Brazil and their potential importance in the arbovirus transmission. A total of 3684 specimens of mosquitoes were captured, 1689 of which caught in the rainy season of 2007, were divided into 78 pools and submitted to viral isolation, Semi-Nested RT-PCR and Nested RT-PCR, with a view to identifying the most important arboviruses in Brazil. Simultaneously, 70 specimens of ticks found blood-feeding on horses were also submitted to the same virological assays. No virus was isolated and viral nucleic-acid detection by RT-PCR was also negative. Nevertheless, a total of 22 Culicidae species were identified, ten of which had previously been reported as vectors of important arboviruses. The diversity of species found blood-feeding on human and horse hosts together with the arboviruses circulation previously reported suggest that the Nhecolândia Sub-region of South Pantanal is an important area for arbovirus surveillance in Brazil.

MOLECULAR CHARACTERIZATION OF INFLUENZA B VIRUS OUTBREAK ON A CRUISE SHIP IN BRAZIL 2012

In February 2012, an outbreak of respiratory illness occurred on the cruise ship MSC Armonia in Brazil. A 31-year-old female crew member was hospitalized with respiratory failure and subsequently died. To study the etiology of the respiratory illness, tissue taken at necropsy from the deceased woman and respiratory specimens from thirteen passengers and crew members with respiratory symptoms were analyzed. Influenza real-time RT-PCR assays were performed, and the full-length hemagglutinin (HA) gene of influenza-positive samples was sequenced. Influenza B virus was detected in samples from seven of the individuals, suggesting that it was the cause of this respiratory illness outbreak. The sequence analysis of the HA gene indicated that the virus was closely related to the B/Brisbane/60/2008-like virus, Victoria lineage, a virus contained in the 2011-12 influenza vaccine for the Southern Hemisphere. Since the recommended composition of the influenza vaccine for use during the 2013 season changed, an intensive surveillance of viruses circulating worldwide is crucial. Molecular analysis is an important tool to characterize the pathogen responsible for an outbreak such as this. In addition, laboratory disease surveillance contributes to the control measures for vaccine-preventable influenza.

SURVIVAL, INDUCTION AND RESUSCITATION OF Vibrio cholerae FROM THE VIABLE BUT NON-CULTURABLE STATE IN THE SOUTHERN CARIBBEAN SEA

The causative agent of cholera, Vibrio cholerae, can enter into a viable but non-culturable (VBNC) state in response to unfavorable conditions. The aim of this study was to evaluate the in situ survival of V. cholerae in an aquatic environment of the Southern Caribbean Sea, and its induction and resuscitation from the VBNC state. V. cholerae non-O1, non-O139 was inoculated into diffusion chambers placed at the Cuare Wildlife Refuge, Venezuela, and monitored for plate, total and viable cells counts. At 119 days of exposure to the environment, the colony count was < 10 CFU/mL and a portion of the bacterial population entered the VBNC state. Additionally, the viability decreased two orders of magnitude and morphological changes occurred from rod to coccoid cells. Among the aquatic environmental variables, the salinity had negative correlation with the colony counts in the dry season. Resuscitation studies showed significant recovery of cell cultivability with spent media addition (p < 0.05). These results suggest that V. cholerae can persist in the VBNC state in this Caribbean environment and revert to a cultivable form under favorable conditions. The VBNC state might represent a critical step in cholera transmission in susceptible areas.

PHLEBOTOMINE FAUNA (DIPTERA: PSYCHODIDAE) IN AN AREA OF FISHING TOURISM IN CENTRAL-WESTERN BRAZIL

The aim of this study was to identify behavioral aspects of the sandfly fauna of a fishing tourism area in the municipality of Bonito (MS). Monthly captures were undertaken from December 2009 to November 2010, using automatic CDC type light traps, from 18h00 to 06h00, in a forested area, a savannah area, peridomiciles and animal shelters near peridomiciliary areas. Nyssomyia whitmani was the most frequent out of a total of 6,699 specimens collected, belonging to 16 species, followed by Psathyromyia bigeniculata and Lutzomyia longipalpis, found in all the environments investigated, though in their greatest numbers in the animal shelters. Ny. whitmani exhibited its highest frequencies during the dry months, coincident with the fishing season, when the risk of transmission of cutaneous leishmaniasis for tourists and inhabitants increases. Noteworthy was the finding of two species naturally infected by flagellates: Ny. whitmani and Pa. bigeniculata. The local population and visiting tourists should be warned of the threat posed by leishmaniasis and the health authorities alerted to the need for adopting environmental sanitary measures, especially regarding such animal shelters as they seem to provide favorable conditions to the proliferation, maintenance and breeding opportunities of phlebotomines.

CHICKEN COOPS, Triatoma dimidiata INFESTATION AND ITS INFECTION WITH Trypanosoma cruzi IN A RURAL VILLAGE OF YUCATAN, MEXICO

This study longitudinally investigated the association between Triatoma dimidiata infestation, triatomine infection with Trypanosoma cruzi and household/backyard environmental characteristics in 101 homesteads in Molas and Yucatan, Mexico, between November 2009 (rainy season) and May 2010 (dry season). Logistic regression models tested the associations between insect infestation/infection and potential household-level risk factors. A total of 200 T. dimidiata were collected from 35.6% of the homesteads, mostly (73%) from the peridomicile. Of all the insects collected, 48% were infected with T. cruzi. Infected insects were collected in 31.6% of the homesteads (54.1% and 45.9% intra- and peridomiciliary, respectively). Approximately 30% of all triatomines collected were found in chicken coops. The presence of a chicken coop in the backyard of a homestead was significantly associated with both the odds of finding T. dimidiata (OR = 4.10, CI 95% = 1.61-10.43, p = 0.003) and the presence of triatomines infected with T. cruzi (OR = 3.37, CI 95% = 1.36-8.33, p = 0.006). The results of this study emphasize the relevance of chicken coops as a putative source of T. dimidiata populations and a potential risk for T. cruzi transmission.

Symptomless Plasmodium vivax parasitemias and malaria eradication in Santa Catarina State, Brazil

Thirty-nine symptomless carriers of P. vivax parasites in the blood gave blood films at monthly intervals for four to six months during the non- transmission season. It was found that parasitaemias can continue for many months. Thirteen of those studied relapsed with symptoms and were treated with chloroquine at a dosage of 600 mg for adults with proportionate doses for children. Of these nine relapsed silently while under observation: a symptomless relapse rate of approximately 70 per cent. One case had symptoms attributable to malaria close to the time of the original survey (the day before). Of the remaining 38 asymptomatic parasite carriers four showed microgametocytes in a density that suggested a potentially high infectivity and six showed microgametocytes in a density suggesting a potentially low to moderate infectivity for mosquito vectors. There was thus a proportion of one smptomatic case of malaria to 10 potentially infective symptomless parasite carriers. Because they feel no need to seek treatment, such persons may form an important reservoir of infection when vectors cannot be fully controlled by spraying. Some possible methods of dealing with such situations are discussed.

Relationship between rainfall and temperature: observations on the cases of visceral leishmaniasis in São Luis Island, State of Maranhão, Brazil

INTRODUCTION: Visceral leishmaniasis is a serious public health problem that requires global control strategies, especially with respect to factors that may intervene in reducing the incidence of endemicity. In this work, rainfall density and temperature were correlated with the incidence of human cases in an area endemic for leishmaniasis in São Luis do Maranhão, Northeastern Brazil. METHODS: Notification of human cases by the National Health Foundation/Regional Coordination of Maranhão (FUNASA/COREMA) from 2002 to 2010 was used. Ecological data (mean temperature and rainfall density) were provided by the Meteorological Office of State. RESULTS: A significant association was verified between the number of VL cases and rainfall rate but not in the analysis concerning mean temperatures. CONCLUSIONS: These data suggest that the control actions in visceral leishmaniasis should be performed during rainy season in the State of Maranhão, which is in the first half of the year.

Occurrence of synanthropic triatomines (Hemiptera: Reduviidae) in the Federal District of Brazil

INTRODUCTION: Entomological surveillance of Chagas disease in the Federal District of Brazil (DF), has recorded the following triatomine species: Panstrongylus megistus, P. geniculatus, P. diasi, Rhodnius neglectus, Triatoma pseudomaculata, and T. sordida. We aimed to analyze the spatial and temporal occurrence of triatomine species collected in DF, and their indices of natural infection with trypanosomes. METHODS: The Health State Secretariat of DF recorded triatomines between 2002 and 2010 in 20 administrative regions. This retrospective analysis considered the number of adults and nymphs of each species collected and infected in both intradomicile and peridomicile. RESULTS: A total of 754 triatomines were collected in 252 reported domiciles. Panstrongylus megistus was the most frequent species (65%), followed by T. pseudomaculata (14%). Of the 309 examined insects, only 3 (1%) specimens of P. megistus were infected with flagellates morphologically similar to Trypanosoma cruzi. The spatial occurrence indicated a higher diversity of triatomines and frequency of T. sordida in rural areas. Moreover, there was a predominance of P. megistus in urban areas. The number of records of P. megistus in the rainy season was two times higher than that during the dry season. The largest number of triatomines was collected in November. CONCLUSIONS: The presence of P. megistus specimens infected with trypanosomes in domiciles, shows the potential risk of human infection in DF. Thus, it is essential to continue entomological surveillance, intensifying it in the rainy season and in regions of greater occurrence.

Analysis of sandflies (Diptera: Psychodidae) in Barra do Garças, State of Mato Grosso, Brazil, and the influence of environmental variables on the vector density of Lutzomyia longipalpis (Lutz & Neiva, 1912)

INTRODUCTION: Leishmaniasis is an infectious and parasitic zoonotic, non-contagious, vector-borne disease caused by protozoa of the genus Leishmania. In Brazil, the major vector of Leishmania (Leishmania) infantum chagasi (Cunha & Chagas, 1934) is Lutzomyia longipalpis. Barra do Garças, State of Mato Grosso, was designated as a priority area by the Brazilian Ministry of Health for american visceral leishmaniasis, and it is important to identify the vector species present in this municipality. Our objective was to raise sandflies and study the influence of environmental variables on the vector density of Lutzomyia longipalpis. METHODS: We performed entomological monitoring in 3 districts using Centers for Disease Control and Prevention (CDC) light traps and recorded human cases of american visceral leishmaniasis in the city. We calculated the relative frequency and richness of sandflies and applied a transfer function model to the vector density correlate with relative humidity. RESULTS: The sandfly population was composed of 2 genera and 27 species, totaling 8,097 individuals. Monitoring identified Lutzomyia longipalpis (44%), followed by Lutzomyia lenti (18.9%), Lutzomyia whitmani (13.9%), Lutzomyia carmelinoi (9.1%), Lutzomyia evandroi (5.1%), Lutzomyia termitophila (3.3%), Lutzomyia sordellii (1.9%), and 20 other species (<4%). The male:female ratio was 3.5:1. We observed high species diversity (Dα = 6.65). Lutzomyia longipalpis showed occurrence peaks during the rainy season; there was a temporal correlation with humidity, but not with frequency or temperature. CONCLUSIONS: The presence of Lutzomyia longipalpis in the urban area of Barra do Garças underscores the changing disease profile, which was previously restricted to the wild environment.

Sandfly fauna in an area endemic for visceral leishmaniasis in Aracaju, State of Sergipe, Northeast Brazil

INTRODUCTION: In recent years, visceral leishmaniasis, a major public health problem, has been spreading from the rural to urban areas in many areas of Brazil, including Aracaju, the capital of the State of Sergipe. However, there are no studies of the sandfly fauna in this municipality or its variation over the year. METHODS: Phlebotomine sandflies were collected from a rural area of Aracaju from September 2007 to July 2009. Modified CDC ultra-violet (UV) light traps were used to evaluate sandfly monthly distribution and their presence in the domestic and peridomestic environments. RESULTS: The most abundant species was Lutzomyia longipalpis (90.4%) followed by Evandromyia lenti (9.6%). A chicken shed trap site had the highest proportion of L. longipalpis (51.1%) and large numbers of L. longipalpis were also collected in the houses closest to the chicken shed. There was a positive correlation between monthly rainfall and L. longipalpis abundance. CONCLUSIONS: Lutzomyia longipalpis is the most abundant species and is probably the main vector of the visceral leishmaniasis agent in the rural area of Aracaju. An increase in L. longipalpis frequency was observed during the rainy season. The peridomicile-intradomicile observations corroborate the importance of chicken sheds for the presence of L. longipalpis in the peridomestic environment. The great numbers of L. longipalpis inside the houses confirm the endophilic behaviour of this species and the possibility of visceral transmission in the intradomicile.

Occurrence of Triatoma costalimai (Hemiptera: Reduviidae) in different environments and climatic seasons: a field study in the Brazilian savanna

INTRODUCTION: Trypanosoma cruzi-infected specimens of Triatoma costalimai have been detected in domiciliary units of Central Brazil, thereby maintaining the potential risk of vectorial transmission of Chagas disease. The aim of this study was to determine the occurrence and natural infection of T. costalimai in different environments (gallery forest, dry forest and peridomicile) and climatic seasons (wet and dry), in the municipality of Mambaí, State of Goiás, Brazil. METHODS: Triatomines were captured in October 2010 and in June 2011, employing two different methods (manual capture and mouse-baited adhesive traps). The insects were later separated by sex and nymphal stage, counted and examined parasitologically by abdominal compression and microscopic analysis of feces. RESULTS: Triatoma costalimai was found in three environments and in the two seasons studied. Overall, capture success of 900 traps and 60 blocks of rocks inspected was 5.8% and 11.7%, respectively. The occurrence of T. costalimai was higher among rocks in the peridomicile, where 97% of the 131 specimens were captured. The proportion of nymphs (98%) was much higher than that of adults, which were only detected in peridomicile. Most (95%) insects were captured during the wet season, with predominance of early-stage nymphs. None of the 43 specimens examined were infected by trypanosomatids. CONCLUSIONS: The results indicate a greater occurrence of T. costalimai in peridomiciliary environments and during the wet season in Mambaí, Goiás, highlighting the synanthropic behavior of this triatomine species in one area of the Brazilian savanna and the importance of entomological surveillance.

Ecology of phlebotomines (Diptera, Psychodidae) in rural foci of leishmaniasis in tropical Brazil

INTRODUCTION: This work aimed to study the community structure of sandflies, with regard to the richness, constancy, abundance, and monthly frequency of the species with a focus on the transmission of leishmaniasis. METHODS: The study was conducted in the rural villages of Bom Jardim and Santa Maria, situated on the edge of a tropical rain forest in the municipality of São Jose de Ribamar, Maranhão, Brazil. The phlebotomines were captured in the intradomiciles and peridomiciles of each village, with Centers for Disease Control (CDC) light traps set in 10 homes in each village, for 1 year, once a month, from 18h to 6h. RESULTS: We collected 1,378 individuals of 16 sandfly species. The capture success rate was higher in Bom Jardim (0.61 specimens/hour/trap) than that of Santa Maria (0.35/specimens/hour/trap). The sandflies were more abundant in the peridomiciles (86.1%) and in the rainy season (77%). Five species were considered constants (occurring in more than 50% of samples), 5 accessory (25%-50%), and 6 accidental (<25%). The most abundant species were Lutzomyia longipalpis (59.7%) and L whitmani (28%). The permutation analysis showed differences between the species composition of the villages and no separation between the intradomicile and peridomicile of each village. The species that most contributed to the dissimilarity between the light traps of the 2 villages were L. longipalpis, L. whitmani, and L. evandroi, contributing to 80.8% of the variation among groups. CONCLUSIONS: The high level of richness and abundance of species and the presence of competent vectors throughout the year and around houses justify the occurrence of leishmaniasis cases reported in the area.

Larval control of Anopheles (Nyssorhinchus) darlingiusing granular formulation of Bacillus sphaericus in abandoned gold-miners excavation pools in the Brazilian Amazon Rainforest

INTRODUCTION: Use of a Bacillus sphaericus based mosquito larvicide was evaluated as an intervention for malaria vector control at a mining site in Amapá, Brazil. Impacts on larval and adult densities of the primary vector Anopheles darlingi were measured over the course of a 52 week study period.METHODS: In Calçoene, State of Amapá, gold mining activity occurs in 19 mining sites in gold-miners of Lourenço. Large pools are formed in mining sites and naturally colonized by Anopheles darlingi. During one year, the impact of applications of VectoLex(r) CG to these larval sources was evaluated. Applications of 20kg/ha were made as needed, based on 10 immature (3rd, 4th instars and pupae) surveillance of health and established thresholds. RESULTS: One hundred percent initial control was observed 48h after each treatment. The pools received from 2-10 (5.3±1.6) treatments during the year. The average re-treatment interval in productive pools was 9.4±4.3 weeks. During weeks 3-52 of the study, mean density of late stage larvae was 78% and pupae were 93% lower in the treated pools than in untreated pools (p< 0.0001, n=51) while reduction of adult mosquitoes was 53% in comparison to the untreated area during the last five months of the study, which were the rainy season (p<0.001).CONCLUSIONS:VectoLex(r) CG reduced immature Anopheles darlingi infestation levels during the entire study period, and reduced adult mosquito populations during the rainy season.

Pediatric epidemiological aspects of scorpionism and report on fatal cases from Tityus stigmurus stings (Scorpiones: Buthidae) in State of Pernambuco, Brazil

Introduction Envenomation by scorpion stings is a major public health problem in numerous tropical countries because of its frequent incidence and potential severity. Approximately 1,900 species of scorpions are known in the world, and at least 130 of these have been described in Brazil. Methods This work reports on 3 child deaths caused by Tityus stigmurus stings and characterizes epidemiological and clinical surveys on pediatric cases of scorpionism recorded in the Centro de Assistência Toxicológica de Pernambuco (Ceatox-PE). Results Scorpion stings accounted for more than 60% of all cases recorded for venomous animals. The children were from 37 cities of the Pernambuco state and accounted for 28.8% of the victims treated for scorpion stings, with the highest incidence in the metropolitan area of Recife. Stings occurred throughout the year and slightly increased during the rainy season. Independent of the elapsed time for a prognosis, most cases showed mild symptoms. Three moderate cases that resulted in death featured cardiogenic shock and/or pulmonary edema or severe neurological symptoms. For the first time, death attributed to T. stigmurus was confirmed by the presence of the scorpion. Conclusions These results suggest that scorpionism in Pernambuco is a public health problem that needs to be monitored carefully throughout the year by the government.