Trophic status of a coastal lagoon - marine harbor system: Potential outwelling rates to the Mesoamerican Barrier Reef southern region.

Eutrophication is still a serious problem in many coastal areas, including the tropics, where river discharges of nutrients is usually high. The ecological stability and ecosystem services of the Mesoamerican Barrier Reef System (MBRS), the world's second-largest coral reef system, suffer a generalized impact by riverine discharge of sediment and organic and inorganic nutrients, which may lead to coastal eutrophication and a coral-macroalgal phase shift. However, few data exist on the MRBS coastal zone status, particularly in Honduras. Here, two in situ sampling campaigns were carried out (May 2017 and January 2018) in the Alvarado Lagoon and Puerto Cortés Bay (Honduras). Measurements included water column nutrients, chlorophyll-a (Chla), particulate organic and inorganic matter and net community metabolism, completed with satellite images analysis. The lagoon and bay environments are ecologically different systems and present different sensitivities to seasonal changes in precipitation as shown by the multivariate analysis. Nonetheless, net community production and respiration rates were neither different spatially, nor seasonally. In addition, both environments were highly eutrophic as shown by the TRIX index. Thus, the Puerto Cortés system represents an important source of dissolved nutrients and particulate matter to the coastal zone. Even though offshore, water quality, based on estimated outwelling rates from the Puerto Cortés system to the coastal waters of the southern MRBS region, improved considerably, concentrations of Chla and nutrients remained higher than those typically measured in non-polluted coral reefs in the Caribbean region and the suggested threshold values. In situ monitoring and assessment of these aspects are crucial to evaluate the ecological functioning of and threats on the MBRS, and elaborate and implement adequate policies for integrated management given its regional and global importance.

Seasonal cycles of phytoplankton biomass and primary production in a tropical temporarily open-closed estuarine lagoon - The effect of an extreme climatic event.

Temporarily open-closed estuaries and estuarine lagoons are among the most complex aquatic ecosystems, prone to undergo rapid changes in response to global change and other anthropogenic impacts. Nonetheless, studies on the factors that control annual cycles of phytoplanktonic biomass and primary production in such systems, especially tropical ones, are still scarce. Even less information exists on the effect increasingly frequent extreme climatic events (ECE) might have on their dynamics. For this purpose, we monitored the changes in ecological conditions in the Los Micos estuarine lagoon (Honduras) by sampling monthly during an annual cycle that included several changes in the lagoon's mouth phase and attempted to understand which environmental factors affect phytoplanktonic biomass and primary production. We also evaluated the impact of, and recovery from, a tropical storm ECE. Annual mean net production (Pn), integrated for the euphotic zone, (4.3 ± 2.8 gC m-2 d-1) and Chlorophyll a (27.1 ± 19.1 mg m-3) values in Los Micos place it as one of the more productive estuaries worldwide. The physico-chemical characteristics of the lagoon clearly depended on mouth phase; however, the values of Chla and Pn did not show significant differences between the open and closed phases. The application of distance-based multivariate linear models did not show any clear dominant model being able to explain the observed Chla and Pn patterns. The most parsimonious models included among others, salinity, particulate organic carbon and PO43-, which suggests that primary production is controlled by multiple factors. During the ECE, about 19% of DIN, 91% of DSi and PO43-, 60% of particulate organic carbon and nitrogen, and 86% of Chla were exported to the sea, greatly reducing Pn. However, Chla and Pn values recovered to pre-storm levels within 30 days, indicating that these biological variables are highly resilient in Los Micos Lagoon.

Method for Malaria Diagnosis Based on Extractions of Samples Using Non-Invasive Techniques: An Opportunity for the Nursing Clinical Practice.

Malaria has been for millennia one of the best known and most destructive diseases affecting humans. Its high impact has aroused great interest for the development of new effective and reliable diagnostic techniques. Recently it has been recently published that hairs from mammal hosts are able to capture, hold and finally remove foreign DNA sequences of Leishmania parasites. The aim of this study was to check if Plasmodium falciparum (P. falciparum) DNA remains stable in blood samples deposited in Whatman paper after suffering different transport and storage conditions, and to compare the sensitivity of these results with those offered by thick a smear and Rapid Diagnostic Test, and besides to examine whether P. falciparum DNA would be detected and quantified by Real time quantitative PCR (qPCR) from hairs of people with different types of malaria. P. falciparum Histidine Repeat Protein II (pHRP-II) antigen detection and P. falciparum DNA were detected in 18 of 19 dry blood samples adhered to Whatman paper (94.74%), besides, Plasmodium DNA was also detected in seven out of 19 hair samples analyzed (36.84%), remaining stable until analysis for several months under the exposure to different environmental conditions. Although the sensitivity of PCR for the diagnosis of malaria in hair samples is not as high as blood analysis, the study of Plasmodium DNA presence in blood and hair could constitute a complementary tool with numerous advantages in sample collection, transport and storage. We suggest that the method could be also applied to medical, forensic and paleo-parasitological diagnosis, not only for malaria but also for searching many other pathogens in hair samples.

Application of qPCR method to hair and cerumen samples for the diagnosis of canine leishmaniosis in Araçatuba, Brazil.

Visceral leishmaniosis (VL) remains a serious public health problem in Brazil. Dogs are the main hosts of the parasite, developing canine leishmaniosis (CanL), hence the importance of an accurate diagnosis of the animals. Recently, the application of qPCR method to non-invasive samples obtained from dogs with CanL has shown high sensitivity. Thus, we analyzed by qPCR blood, hair (from healthy zones and cutaneous lesions) and cerumen of 16 dogs with confirmed leishmaniosis from Araçatuba, a Brazilian endemic area. Cerumen-qPCR showed the highest sensitivity (87.5%), followed by hair (lesions: 78.57%, healthy skin: 62.5%), and blood (68.75%). We also analyzed blood, hair and cerumen of 5 healthy dogs from a non-endemic area, obtaining 100% of specificity in all samples. The use of cerumen and hair for qPCR analysis provides high reliability, taking into account the sensitivity and total specificity of the method. The non-invasive sampling procedure without the need of specific conditions of storage and transport support the usefulness of hair and cerumen for the diagnosis of CanL.

First detection of Leishmania kDNA in canine cerumen samples by qPCR.

Nowadays, searching for alternative non-invasive methods for molecular diagnosis of canine visceral leishmaniosis is getting increasingly important. We previously described the presence of Leishmania kinetoplast DNA (kDNA) in canine hair; in this case we hypothesized whether foreign DNA might be present in cerumen of dogs with leishmaniosis, and be detected by Real time quantitative PCR (qPCR). A population of 38 dogs that lived in Leishmania endemic areas was divided in two groups: A (33 dogs with confirmed leishmaniosis by serological techniques) and B (5 healthy dogs). Blood, lymph node, bone marrow and cerumen samples from all animals were tested for the presence of parasite kDNA. Our method was 100% specific, and in dogs from group A, Leishmania infantum kDNA was detected and quantified in the 100% of lymph node samples, in 90.9% of cerumen samples, in 88.5% of the bone marrow samples and in 57.6% of the blood samples. The qPCR-cerumen is a new non-invasive method that shows a high potential for the diagnosis of zoonotic visceral leishmaniosis.

Population structure in the endangered Blanca Cacereña bovine breed demonstrated by RAPD analyses.

RAPD analyses have been used to determine the genetic diversity and the population structure of the endangered Blanca Cacereña bovine breed. Genetic variability was evaluated on the basis of 1048 loci produced by 71 primers. RAPD produced a number of polymorphic loci (30.44%), and it has been proved to be a useful method for evaluating polymorphisms in this breed. The dendrograms based on simililarity indexes and on Nei's genetic distances between 60 animals and the value of genetic differentiation among subpopulations (F(ST)) showed a clear population substructure defined by herds and a scarce genetic flow among herds. Analysis of molecular variance (AMOVA) showed that 32.4% of the total variance was due to differences among herds and confirmed the clustering found. The results of the present study allow us to plan more adequate mating in order to maintain the genetic diversity and to improve the efficiency of conservation for the Blanca Cacereña bovine breed.

Detection of Leishmania infantum kinetoplast minicircle DNA by Real Time PCR in hair of dogs with leishmaniosis.

It is known that hair can accumulate environmental toxics and excrete foreign chemical or biological substances. In this context, we hypothesized that foreign DNA could be found in the hair of an infected organism, and thus, be detected by Real Time PCR in the hair of Leishmania infantum naturally infected dogs. A population of 28 dogs living in Leishmania endemic areas was divided into two groups: A (13 Leishmania infected dogs) and B (15 healthy dogs). Blood, lymph node and ear hair samples from all of them were tested for the presence of parasite kinetoplast DNA (kDNA). For the same purpose, hair of several body areas and hair sections of two infected dogs were also analyzed. Epidermal keratinocytes from an infected animal were also analyzed for reactivity against Leishmania antigens by ELISA and for the presence of kDNA. Regarding to dogs from group A, parasite kDNA was detected in the 100% of lymph node samples. The sensitivity of Real Time PCR in ear hair was similar to that obtained in blood (9 positive out of 13 versus 8 positive out of 13, respectively). Moreover, the presence of L. infantum kDNA was also detected in the hair of all the analyzed body zones, in all hair sections and in epidermal keratinocytes. In infected dogs, parasite kDNA could be detected and quantified from just one single hair, whereas it was not detected in any of the samples of the healthy dogs. This work describes a new method for a reliable and non-invasive diagnosis of canine leishmaniosis using hair samples of infected animals. The data presented also provide some insights for the understanding of the physiology of keratinocytes and the role of hair as a specialized tissue in the kidnapping and removal of foreign DNA.

First detection of Leishmania infantum kinetoplast DNA in hair of wild mammals: application of qPCR method to determine potential parasite reservoirs.

The data presented in this paper describe the application of a method for a reliable and non-invasive diagnosis of leishmaniosis in wild reservoirs, based on the detection of Leishmania infantum kinetoplast DNA (kDNA) in hair samples by Real Time PCR (qPCR). The study has been performed on 68 ear/leg hair samples from 5 different wild species (Vulpes vulpes, Canis lupus, Martes foina, Rattus norvegicus and Erinaceus europaeus) from several geographic areas of West and North Spain. The presence of Leishmania kDNA was detected in 14 of the 68 analyzed samples, being the highest quantity of DNA observed in foxes. This is the first report of the presence of Leishmania in a hedgehog. The kDNA remained stable under the exposure of hair to different environmental conditions (freezing or high temperature, ultraviolet rays or treatment with tanning salts). This detection method could constitute a suitable alternative for the search of the parasite in wild hosts, due to the numerous advantages that hair samples present for collection, transport and storage processes.

Detection and chronology of parasitic kinetoplast DNA presence in hair of experimental Leishmania major infected BALB/c mice by Real Time PCR.

Hair can accumulate foreign chemical or biological substances. Recently, it has been reported that parasite DNA can also be detected in the hair of Leishmania infantum infected dogs. The aim of this work has been to find out whether parasite DNA incorporates in the hair of Leishmania major experimentally infected animals. For this purpose, a group of 4 BALB/c mice, intradermally inoculated in both ears with 1000 L. major V1 strain promastigote forms, was monitored for parameters associated to the infection during 35 days. Weekly, ear swelling was measured, and hair samples from ears and leg were collected. Blood samples were obtained before challenge and at day 35 post infection, when parasite load was measured in ear, lymph node and spleen by limit dilution. Ear swelling and other parameters observed in the infected mice were consistent with those described for this model. The presence of parasite kinetoplast DNA (kDNA) was detected by Real Time PCR in all ear and leg hair samples at the final timepoint. These data suggests that hair is a specialized tissue in the sequestration and removal of foreign DNA. Detection of DNA in hair could be, therefore, a useful tool to chronologically record the infection process during experimental mice assays.

Leishmania major infection in susceptible and resistant mice elicit a differential humoral response against a total soluble fraction and defined recombinant antigens of the parasite.

In the present work, we analyzed the humoral response of Leishmania major experimentally infected BALB/c and C57BL/6 mice against three Leishmania antigens: total soluble antigen (soluble leishmania antigen(SLA)), a chimerical recombinant protein formed by the genetic fusion of four cytoplasmic proteins (PQ), and a kinetoplastic membrane protein (Kmp-11). We determined the correlation between the immune response against these proteins and the histopathological changes induced in the susceptible and resistant mice after infection. The data showed the existence of wide differences in the recognition of SLA, PQ, and Kmp-11 by the sera from both strains. The anti-SLA titer of BALB/c was 100 times higher than that of C57BL/6 mice. Antibodies against the recombinant Kmp-11 were detected only in infected BALB/c during the first stage of the infection. In contrast, the PQ protein was recognized by the sera from infected BALB/c mice but exclusively when they were in a late-lesion period. The data suggest that the response against the membrane Kmp-11 protein is transient and correlates with early developmental stages of the infection, whereas the response against cytoplasmic proteins as those present in PQ is sustained and could be considered as a marker of an advanced stage of the infection and disease.