Assessment of PCR in the detection of Leishmania spp in experimentally infected individual phlebotomine sandflies (Diptera: Psychodidae: Phlebotominae)

DNA amplification by the polymerase chain reaction (PCR) was applied in the investigation of the presence of Leishmania (Kinetoplastida: Trypanosomatidae) parasites in single phlebotomine sandflies. Three phlebotomine/parasite pairs were used: Lutzomyia longipalpis/Leishmania chagasi, Lutzomyia migonei/Leishmania amazonensis and Lutzomyia migonei/Leishmania braziliensis, all of them incriminated in the transmission of visceral or cutaneous leishmaniasis. DNA extraction was performed with whole insects, with no need of previous digestive tract dissection or pooling specimens. The presence of either mouse blood in the digestive tract of the sandflies or the digestive tract itself did not interfere in the PCR. Infection by as few as 10 Leishmania sp. per individual were sufficient for DNA amplification with genus-specific primers. Using primers for L. braziliensis and L. mexicana complexes, respectively, it was possible to discriminate between L. braziliensis and L. amazonensis in experimentally infected vectors (L. migonei).Neste trabalho avaliamos o uso da reação em cadeia da polimerase (PCR) na investigação da presença de parasitas Leishmania (Kinetoplastida:Trypanosomatidae) em flebotomíneos individuais. Para isso, foram utilizados três pares flebotomíneo/parasita: Lutzomyia longipalpis/Leishmania chagasi, Lutzomyia migonei/Leishmania amazonensis e Lutzomyia migonei/Leishmania braziliensis, todos eles incriminados na transmissão de leishmaniose cutânea ou visceral. O DNA total a ser amplificado foi extraído de flebotomíneos inteiros, contendo ou não o parasita, sem dissecção prévia do trato digestivo ou combinação de indivíduos. Conteúdos do trato digestivo de flebotomíneos, em especial sangue de camundongo, não interferiram na reação de amplificação. Dez parasitas Leishmania sp. por flebotomíneo foram suficientes para detecção com iniciadores gênero-específicos. Com a utilização de iniciadores para os complexos L. braziliensis e L. mexicana, respectivamente, foi possível discriminar entre L. braziliensis e L. amazonensis, em flebotomíneos infectados experimentalmente (L. migonei)

Carbohydrate and protein metabolism of marandu grass affected by nitrogen fertilisation and number of cuts

Received: June 18th, 2023 ; Accepted: September 30th, 2023 ; Published: October 23rd, 2023 ; Correspondence: [email protected] the metabolism of tropical grasses in response to management practises imposed in pastoral environments allows for improvements in the management and use of mineral fertilisers. This study aimed to quantify metabolite content in different plant parts of Marandu grass (Urochloa brizantha), with a specific focus on the influence of nitrogen fertilisation and its effects following successive cuts. The treatments corresponded to four nitrogen (N) rates (0, 75, 150, and 225 kg N ha-1 ) and the number of cuts (one, two and three cuts). The plants were fractionated into leaves, stems, and roots to assess the content of water-soluble carbohydrates (WSC), starch, albumin, globulin, prolamin, and glutelin content. N fertilisation influenced the WSC and starch content in different parts of the plant, varying according to the cuts made. In the leaves and roots, fertilisation reduced the content of WSC and starch with one cut, as these were utilised as energy sources for assimilating the excess nitrogen in the soil. There was an increase in the concentration of all protein groups with nitrogen fertilisation in all parts of the plant with one cut. In plants cut two and three times, N fertilisation led to specific increases and decreases in different parts of the plants as an adaptive strategy for allocating resources as the number of cuts increased. Our results broaden our understanding of carbohydrate and protein metabolism in tropical grasses, thereby providing subsidies for the rational use of nitrogen fertilisers

MutSα expression predicts a lower disease-free survival in malignant salivary gland tumors: an immunohistochemical study

Background: Appropriate DNA replication is vital to maintain cell integrity at the genomic level. Malfunction on DNA repair mechanisms can have implications related to tumor behavior. Our aim was to evaluate the expression of key complexes of the DNA mismatch-repair system MutSα (hMSH2-hMSH6) and MutSβ (hMSH2-hMSH3) in a panel comprising the most common benign and malignant salivary gland tumors (SGT), and to determine their association with disease-free survival. Material and Methods: Ten cases of normal salivary gland (NSG) and 92 of SGT (54 benign and 38 malignant) were retrieved. Immunohistochemistry was performed for hMSH2, hMSH3, hMSH6. Scanned slides were digitally analyzed based on the percentage of positive cells with nuclear staining. Cases were further classified in MutSαhigh and MutSβhigh based on hMSH2-hMSH6 and hMSH3-hMSH6 expression, respectively. Results: hMSH3 expression was lower in malignant SGT compared to NSG and benign cases. Adenoid cystic carcinoma (ACC) cases with perineural invasion presented a lower percentage of hMSH3 positive cells. hMSH6 was downregulated in both benign and malignant SGT compared to NSG. Malignant SGT cases with MutSαhigh expression had lower disease-free survival compared to MutSαlow cases. A 10.26-fold increased risk of presenting local recurrence was observed. Conclusions: Our findings suggest that a lack of hMSH3 protein function is associated with a more aggressive phenotype (malignancy and perineural invasion) and that MutSα overexpression predicts a poor clinical outcome in malignant SGT

Epidemiological situation of bovine brucellosis in the State of São Paulo, Brazil

Realizou-se um estudo para caracterizar a situação epidemiológica da brucelose bovina no Estado de São Paulo. O Estado foi estratificado em sete circuitos produtores de bovinos, e 150 rebanhos com atividade reprodutiva foram aleatoriamente selecionados em cada um deles. No total, foram amostrados 1.073 rebanhos. Foram aletoriamente coletadas amostras de soro de 10 ou 15 fêmeas bovinas com idade ³24 meses, totalizando 8.761 animais. Os soros foram submetidos a um protocolo de testes em série, tendo o teste do antígeno acidificado tamponado como método de triagem e o da fixação de complemento como confirmatório. A prevalência estimada de rebanhos com pelo menos um animal soropositivo foi de 9,7% [7,8-11,6%], enquanto a prevalência estimada de animais soropositivos foi de 3,8% [0,7-6,9%], no Estado. Em cada rebanho foi aplicado um questionário epidemiológico para avaliar o grau de associação de possíveis fatores de risco (odds ratio, OR) com a doença. Propriedades com 87 ou mais bovinos (OR= 2,25) e compra de reprodutores (OR= 1,56) foram as variáveis mais associadas à condição de foco de brucelose. ____________________________________________________________________________________________________________ ARQUIVOA study to characterize the epidemiological situation of bovine brucellosis in the State of São Paulo was carried out. The state was stratified in seven cattle production regions and 150 herds with reproductive activity were randomly selected within each one. A total of 1,073 herds were sampled. In a second stage, 10 or 15 cows older than 24 month age (in herds with <100 cows and herds ≥100 cows, respectively) were bled at random. A total of 8,761 animals were bled. Sera were tested using a serial testing procedure with the Rose Bengal test as the screening method and the Complement Fixation as the confirmatory method. The estimated prevalence of herds with at least one positive animal was 9.7% [7.8-11.6%], whereas the estimated prevalence of positive cattle was 3.8% [0.7-6.9%] in São Paulo State. An epidemiological questionnaire was applied in each farm, allowing the evaluation of the association of selected risk factors with the disease. The characteristics associated with the brucellosis infected herds were farms with more than 87 bovines (OR= 2.25) and introduction of breeding cattle (OR= 1.56)

Epidemiological situation of bovine brucellosis in the State of Rio Grande do Sul, Brazil

Realizou-se um estudo para caracterizar a situação epidemiológica da brucelose bovina. O Estado foi dividido em sete regiões. Em cada região foram amostradas aleatoriamente cerca de 300 propriedades, e dentro dessas foi escolhido de forma aleatória um número pré-estabelecido de animais, dos quais foi obtida uma amostra de sangue. No total foram amostrados 16.072 animais, provenientes de 1.957 propriedades. Em cada propriedade amostrada foi aplicado um questionário epidemiológico para verificar o tipo de exploração e as práticas zootécnicas e sanitárias que poderiam estar associadas ao risco de infecção pela doença. O protocolo de testes utilizado foi o da triagem com o teste do antígeno acidificado tamponado e o reteste dos positivos com o teste do 2-mercaptoetanol. O rebanho foi considerado positivo se pelo menos um animal foi reagente às duas provas sorológicas. Para o Estado, as prevalências de focos e de animais infectados foram, respectivamente, 2,1% [1,5-2,6%] e 1,0% [0,60-1,4%]. Para os circuitos, a prevalência de focos e a de animais foram, respectivamente: circuito 1, 3,1% [1,4-5,7%] e 0,95% [0,0-2,0%]; circuito 2, 7,7% [4,9-11,3%] e 1,0% [0,40-1,7%]; circuito 3, 5,7% [3,4-8,8%] e 2,1% [0,41-3,8%]; circuito 4, 0,66% [0,08-2,4%] e 0,66% [0,0-1,8%]; circuito 5, 0,66% [0,08-2,4%] e 0,05% [0,0-0,13%]; circuito 6, 0,0% [0,0-1,3%] e 0,0% [0,0-0,25%]; circuito 7, 5,4% [2,5-10,1%] e 2,9% [0,49-5,3%]. Os fatores de risco (odds ratio, OR) associados à condição de foco foram: exploração de corte (OR= 4,27 [1,82-10,01]) e histórico de aborto (OR=3,27,1,71-6,25]). ____________________________________________________________________________________________________________ ABSTRACTA study to characterize the epidemiological status of bovine brucellosis was carried out in the State of Rio Grande do Sul. The State was divided in seven regions. Three hundred herds were randomly sampled in each region and a pre-established number of animals were sampled in each of these herds. A total of 16,072 serum samples from 1,957 herds, were collected. In each herd, it was applied an epidemiological questionnaire focused on herd traits as well as husbandry and sanitary practices that could be associated with the risk of infection. The serum samples were screened for antibodies against Brucella spp. by the Rose-Bengal Test and all positive sera were re-tested by the 2-mercaptoethanol test. The herd was considered positive if at least one animal was positive on both tests. The prevalences of infected herds and animals in the State were, respectively 2.1% [1.5-2.6%] and 1.0% [0.60-1.4%]. In the regions, the prevalences of infected herds and animals were, respectively: region 1, 3.1% [1.4-5.7%] and 0.95% [0.0-2.0%]; region 2, 7.7% [4.9-11.3%] and 1.0% [0.40-1.7%]; region 3, 5.7% [3.4-8.8%] and 2.1% [0.41-3.8%]; region 4, 0.66% [0.08-2.4%] and 0.66% [0.0-1.8%]; region 5, 0.66% [0.08-2.4%] and 0.05% [0.0-0.13%]; region 6, 0.0% [0.0-1.3%] and 0.0% [0.0-0.25%]; and region 7, 5.4% [2.5-10.1%] and 2.9% [0.49-5.3%]. The risk factors (odds ratio, OR) associated with the presence of infection were: beef herd (OR= 4.27 [1.82-10.01]) and recent history of abortion (OR= 3.27-1.71-6.25])

Near-intrinsic energy resolution for 30-662 keV gamma rays in a high pressure xenon electroluminescent TPC

We present the design, data and results from the NEXT prototype for Double Beta and Dark Matter (NEXT-DBDM) detector, a high-pressure gaseous natural xenon electroluminescent time projection chamber (TPC) that was built at the Lawrence Berkeley National Laboratory. It is a prototype of the planned NEXT-100 136Xe neutrino-less double beta decay (0νββ) experiment with the main objectives of demonstrating near-intrinsic energy resolution at energies up to 662 keV and of optimizing the NEXT-100 detector design and operating parameters. Energy resolutions of ∼1% FWHM for 662 keV gamma rays were obtained at 10 and 15 atm and ∼5% FWHM for 30 keV fluorescence xenon X-rays. These results demonstrate that 0.5% FWHM resolutions for the 2,459 keV hypothetical neutrino-less double beta decay peak are realizable. This energy resolution is a factor 7 to 20 better than that of the current leading 0νββ experiments using liquid xenon and thus represents a significant advancement. We present also first results from a track imaging system consisting of 64 silicon photo-multipliers recently installed in NEXT-DBDM that, along with the excellent energy resolution, demonstrates the key functionalities required for the NEXT-100 0νββ search