Detection and molecular characterization of Cryptosporidium spp. in captive canaries (Serinus canaria) using different diagnostic methods

Abstract This study used several diagnostic methods to examine the occurrence of and molecularly characterize Cryptosporidium spp. in captive canaries (Serinus canaria) in southern and southeastern Brazil. A total of 498 fecal samples were purified by centrifugal-flotation using Sheather's solution. Cryptosporidium spp. diagnosis was performed using three diagnostic methods: malachite green negative staining, nested PCR targeting the 18S rRNA gene, followed by sequencing the amplified fragments, and duplex real-time PCR targeting the 18S rRNA specific to detect Cryptosporidium galli and Cryptosporidium avian genotype III. The overall positivity for Cryptosporidium spp. (total samples positive in at least one protocol) from the microscopic analysis, nested PCR and duplex real-time PCR protocol results was 13.3% (66/498). The positivity rates were 2.0% (10/498) and 4.6% (23/498) for Cryptosporidium spp. by microscopy and nested PCR, respectively. Sequencing of 20 samples amplified by nested PCR identified C. galli (3.0%; 15/498), Cryptosporidium avian genotype I (0.8%; 4/498) and Cryptosporidium avium (0.2%; 1/498). Duplex real-time PCR revealed a positivity of 7.8% (39/498) for C. galli and 2.4% (12/498) for avian genotype III. Malachite green negative staining differed significantly from nested PCR in detecting Cryptosporidium spp. Duplex real-time PCR was more sensitive than nested PCR/sequencing for detecting gastric Cryptosporidium in canaries

Strategies to minimize heat stress in broilers chickens

The objective of this study was to evaluate the application of early age thermal conditioning (TC) and dietary electrolyte balances (DEB) to minimize the effect of heat stress. A total of 240 Cobb 500 1-d-old male broiler chicks was randomly allocated to 24 floor pens with six replicates per treatment in a 2x2 factorial arrangement (with and without TC and with and without DEB). Dietary treatments consisted: (T1) a traditional diet without TC; (T2) traditional diet with TC; (T3) with the application of dietary electrolyte and without TC and (T4) application of dietary electrolyte with TC. The thermal conditioning was conducted at 5 d of age (36°C for 24 h), only half of the batch (120 birds). After this period, all birds were transferred to boxes of 1.5 x 3m (10 birds / box), with wood shavings reused as litter. Chicks were exposed to chronic heat stress (32°C) for 6 h from 35 to 39 d of age, in all treatments, being electronically monitored the temperature and humidity of the microclimate of the birds. Feed and water were provided ad libitum, even during periods of stress. Were measured performance data (weight gain, feed intake and feed conversion) and mortality rate. Data were subjected to ANOVA using the GLM procedures of SAS. The results of this study demonstrated no interaction effects of all evaluated parameters (performance and mortality). Therefore, no synergism occurred when both strategies (TC and DEB) were applied. Only the first week there was prejudice on broiler performance with the application of the TC and apparently the technique of early age thermal conditioning no improve the resistance of broilers to chronic heat stress. On the other hand, DEB had significantly favorable effects (P<0.05) on performance and minimized mortality immediately after application of the TC

Prevalence of ehrlichia canis (rickettsiales: ehrlichieae) DNA in tissues from rhipicephalus sanguineus (acari: ixodidae) ticks in areas endemic for canine monocytic ehrlichiosis in Brazil

Canine monocytic ehrlichiosis (CME) is a disease caused by the obligate intracellular bacterium Ehrlichia canis. Tropical lineages of Rhipicephalus sanguineus ticks play an essential role in the transmission of this pathogen. The aim of the present study was to evaluate the prevalence of E. canis DNA in tissue from R. sanguineus ticks in areas endemic for CME in Brazil and quantify levels of E. canis DNA in dissected tissues from these samples. A total of 720 ticks were collected from 72 dogs (36 dogs from the city Aracatuba in Sao Paulo state and 36 from Campo Grande in the state of Mato Grosso do Sul). Ticks were dissected to collect the guts, ovaries and salivary gland. A quantitative polymerase chain reaction (qPCR) targeting the disulphide bond formation (dsb) protein gene was performed to quantify the level of E. canis infection. The E. canis dsb-qPCR assay was positive for 31.9, 10, and 15.2% of the gut, ovary, and salivary glands, respectively. The average gut, ovary, and salivary gland bacterial load estimated by qPCR was 1.21 x 10(3), 2.60 x 10(3), and 4.92 x 10(3) gene copies/mu l, respectively. This is the first report of E. canis DNA in ovaries of R. sanguineus ticks parasitizing dogs in these CME-endemic areas. These observations raise the possibility of E. canis trans-ovarial transmission563828831FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP2014/26461-

Detection and molecular characterization of Cryptosporidium spp. in captive canaries (Serinus canaria) using different diagnostic methods

<div><p>Abstract This study used several diagnostic methods to examine the occurrence of and molecularly characterize Cryptosporidium spp. in captive canaries (Serinus canaria) in southern and southeastern Brazil. A total of 498 fecal samples were purified by centrifugal-flotation using Sheather's solution. Cryptosporidium spp. diagnosis was performed using three diagnostic methods: malachite green negative staining, nested PCR targeting the 18S rRNA gene, followed by sequencing the amplified fragments, and duplex real-time PCR targeting the 18S rRNA specific to detect Cryptosporidium galli and Cryptosporidium avian genotype III. The overall positivity for Cryptosporidium spp. (total samples positive in at least one protocol) from the microscopic analysis, nested PCR and duplex real-time PCR protocol results was 13.3% (66/498). The positivity rates were 2.0% (10/498) and 4.6% (23/498) for Cryptosporidium spp. by microscopy and nested PCR, respectively. Sequencing of 20 samples amplified by nested PCR identified C. galli (3.0%; 15/498), Cryptosporidium avian genotype I (0.8%; 4/498) and Cryptosporidium avium (0.2%; 1/498). Duplex real-time PCR revealed a positivity of 7.8% (39/498) for C. galli and 2.4% (12/498) for avian genotype III. Malachite green negative staining differed significantly from nested PCR in detecting Cryptosporidium spp. Duplex real-time PCR was more sensitive than nested PCR/sequencing for detecting gastric Cryptosporidium in canaries.</p></div