The epidemiology of canine leishmaniasis: transmission rates estimated from a cohort study in Amazonian Brazil

We estimate the incidence rate, serological conversion rate and basic case reproduction number (R0) of Leishmania infantum from a cohort study of 126 domestic dogs exposed to natural infection rates over 2 years on Marajó Island, Pará State, Brazil. The analysis includes new methods for (1) determining the number of seropositives in cross-sectional serological data, (2) identifying seroconversions in longitudinal studies, based on both the number of antibody units and their rate of change through time, (3) estimating incidence and serological pre-patent periods and (4) calculating R0 for a potentially fatal, vector-borne disease under seasonal transmission. Longitudinal and cross-sectional serological (ELISA) analyses gave similar estimates of the proportion of dogs positive. However, longitudinal analysis allowed the calculation of pre-patent periods, and hence the more accurate estimation of incidence: an infection–conversion model fitted by maximum likelihood to serological data yielded seasonally varying per capita incidence rates with a mean of 8·66×10[minus sign]3/day (mean time to infection 115 days, 95% C.L. 107–126 days), and a median pre-patent period of 94 (95% C.L. 82–111) days. These results were used in conjunction with theory and dog demographic data to estimate the basic reproduction number, R0, as 5·9 (95% C.L. 4·4–7·4). R0 is a determinant of the scale of the leishmaniasis control problem, and we comment on the options for control

Infectiousness in a Cohort of Brazilian Dogs: Why Culling Fails to Control Visceral Leishmaniasis in Areas of High Transmission

The elimination of seropositive dogs in Brazil has been used to control zoonotic visceral leishmaniasis but with little success. To elucidate the reasons for this, the infectiousness of 50 sentinel dogs exposed to natural Leishmania chagasi infection was assessed through time by xenodiagnosis with the sandfly vector, Lutzomyia longipalpis. Eighteen (43%) of 42 infected dogs became infectious after a median of 333 days in the field (105 days after seroconversion). Seven highly infectious dogs (17%) accounted for >80% of sandfly infections. There were positive correlations between infectiousness and anti-Leishmania immunoglobulin G, parasite detection by polymerase chain reaction, and clinical disease (logistic regression, r2 = 0.080.18). The sensitivity of enzyme-linked immunosorbent assay to detect currently infectious dogs was high (96%) but lower in the latent period (<63%), and specificity was low (24%). Mathematical modeling suggests that culling programs fail because of high incidence of infection and infectiousness, the insensitivity of the diagnostic test to detect infectious dogs, and time delays between diagnosis and culling

Tissue Cytokine Responses in Canine Visceral Leishmaniasis

To elucidate the local tissue cytokine response of dogs infected with Leishmania chagasi, cytokine mRNA levels were measured in bone marrow aspirates from 27 naturally infected dogs from Brazil and were compared with those from 5 uninfected control animals. Interferon-γ mRNA accumulation was enhanced in infected dogs and was positively correlated with humoral (IgG1) but not with lymphoproliferative responses to Leishmania antigen in infected dogs. Increased accumulation of mRNA for interleukin (IL)4, IL-10, and IL-18 was not observed in infected dogs, and mRNA for these cytokines did not correlate with antibody or proliferative responses. However, infected dogs with detectable IL-4 mRNA had significantly more severe symptoms. IL-13 mRNA was not detectable in either control or infected dogs. These data suggest that clinical symptoms are not due to a deficiency in interferon-γ production. However, in contrast to its role in human visceral leishmaniasis, IL-10 may not play a key immunosuppressive role in dogs

Detection of Leishmania infantum by PCR, serology and cellular immune response in a cohort study of Brazilian dogs

The sensitivity and specificity of PCR, serology (ELISA) and lymphoproliferative response to Leishmania antigen for the detection of Leishmania infantum infection were evaluated in a cohort of 126 dogs exposed to natural infection in Brazil. For PCR, Leishmania DNA from bone-marrow was amplified with both minicircle and ribosomal primers. The infection status and time of infection of each dog were estimated from longitudinal data. The sensitivity of PCR in parasite-positive samples was 98%. However, the overall sensitivity of PCR in post-infection samples, from dogs with confirmed infection, was only 68%. The sensitivity of PCR varied during the course of infection, being highest (78–88%) 0–135 days post-infection and declining to around 50% after 300 days. The sensitivity of PCR also varied between dogs, and was highest in sick dogs. The sensitivity of serology was similar in parasite-positive (84%), PCR-positive (86%) and post-infection (88%) samples. The sensitivity of serology varied during the course of infection, being lowest at the time of infection and high (93–100%) thereafter. Problems in determining the specificity of serology are discussed. The sensitivity and specificity of cellular responsiveness were low. These data suggest that PCR is most useful in detecting active or symptomatic infection, and that serology can be a more sensitive technique for the detection of all infected dogs

Structural and productive characteristics of Marandu and Xaraés grasses fertilized at different times after harvesting

The objective of this study was to assess the effect of applying N and K2O at different times after harvest on the structure and production characteristics of Marandu and Xaraés cultivars of Brachiaria brizantha. Two greenhouse 4 × 2 factorial experiments using a randomized block design with eight treatments and four replicates were carried out. In experiment 1, the plants in pots were fertilized weekly with 50 mg/dm³ of N and K2O until a week before a uniformity cut, for a total of four applications. After cutting, plants were fertilized with 50 mg/dm³ of N and K2O at 0, 3, 6, and 9 days. Cultivars were evaluated after 44 days of regrowth. In experiment 2, a single 50 mg/dm³ dose of N and K2O was applied a week before the uniformity cut, and 100 mg/dm³ of N and K2O were applied at 0, 4, 8, and 12 days after cutting; cultivars were evaluated after 39 days of regrowth. In experiment 1, the leaf/stem ratio and dry matter yield were influenced by the time at which fertilizers were applied. In the second experiment, the total number of leaves, expanded leaves, dead leaves, and the leaf lamina accumulated length and dry matter production were influenced by the time at which fertilizers were applied. Marandugrass had more leaves and tillers than cv. Xaraés, which, in turn, had greater height, dry matter yield and regrowth vigor. Applying nitrogen and potassium fertilizer immediately after cutting improves the structural characteristics, while the dry matter yield is maximized by applying fertilizers immediately after cutting by using lower doses of N, or at 4.5 days after cutting by using higher doses of N

Crescimento de folhas do capim-bermuda tifton 85 submetido à adubação nitrogenada após o corte Leaf growth of tifton 85 bermudagrass submitted to nitrogen fertilization after cutting

Com o objetivo de avaliar a influência de doses e épocas de aplicação de nitrogênio após o corte no fator de correção de área foliar, na taxa de alongamento de folhas e no comprimento e área da lâmina foliar do capim-bermuda tifton 85 (Cynodon spp) foram conduzidos dois experimentos em casa de vegetação. Ambos os experimentos foram estabelecidos em vasos com capacidade para 7 kg de terra, com solo classificado como Neossolo Quartzarênico Órtico típico, em esquema fatorial 4 × 2, para avaliação de quatro doses de nitrogênio (0, 80, 160 e 240 mg kg-1 de solo) e duas épocas de aplicação (imediatamente após o corte e sete dias após o corte das plantas). O delineamento experimental foi em blocos completos ao acaso, com quatro repetições. Com o fornecimento de nitrogênio, observou-se diminuição no fator de correção de área foliar. O nitrogênio proporciona variação positiva no comprimento foliar, na área da lâmina foliar e na taxa de alongamento da folha, variações que ocorreram em maior grandeza entre a não-aplicação de nitrogênio e a dose de 80 mg kg-1 de solo.<br>With the objective of evaluating the influence of nitrogen rates and application time after cutting on correction factor for leaf area, on leaf elongation rate and on blade leaf length and area of tifton 85 bermudagrass (Cynodon spp), two experiments were carried out in a greenhouse. Both experiments were established in pots with capacity for 7 kg of soil classified as Entisol, in a 4 × 2 factorial scheme, for evaluation of four nitrogen rates (0, 80, 160 and 240 mg kg-1 of soil) and two application times (immediately after cutting and seven days after cutting of the plants). It was used a complete randomized block design, with four replications. As nitrogen was supplied, it was observed a decrease in the correction factor for leaf area. There is a predominance of positive effects of nitrogen on leaf length, on leaf blade area and on leaf elongation rate. Great nitrogen effect was observed when comparying no-nitrogen application with the rate of 80 mg kg-1 of soil