Feeding laying hens with lactobacilli improves internal egg quality and animal health.

Feeding animals with lactobacilli strains is a biotechnological strategy to improve production, food quality, and animal health. Thus, this study aimed to select new lactic acid bacteria (LAB) able to improve laying hens health and egg production. Forty Bovans White layers (two days old) were randomly divided into four experimental groups that receive an oral gavage with saline solution (control group) or with one of the three lactobacilli selected (KEG3, TBB10, and KMG127) by their antagonistic activity against the foodborne pathogen Bacillus cereus GGD_EGG01. 16 S rRNA sequencing identified KEG3 as Lentilactobacillus sp., and TBB10 and KMG127 as Lactiplantibacillus sp. The data showed that feeding birds with LAB increased weight uniformity and improved the internal quality of the eggs (high yolk index and Haugh unit) compared with the control group (p < 0.05). Beta-diversity analysis showed that LAB supplementation modifies the cecal microbiota of laying hens. The prokaryotic families Bacteroidaceae, Ruminococcaceae, Rikenellaceae, and Lactobacillaceae were most important to the total dissimilarity of the cecal microbial community (calculated by SIMPER test). At end of in vivo experiments, it was possible to conclude that the feed of laying hens with Lentilactobacillus sp. TBB10 and Lentilactobacillus sp. KEG3 can be an important biotechnological tool for improving food quality and animal health.

Contaminated water confirmed as source of infection by bioassay in an outbreak of toxoplasmosis in South Brazil.

The protozoan Toxoplasma gondii is a causative agent of toxoplasmosis, an important and widespread zoonotic disease. The transmission of this disease in humans includes ingestion of sporulated oocysts present in contaminated water or food. T. gondii oocysts are widely distributed and toxoplasmosis is considered a major food- and waterborne pathogen worldwide, making drinking water containing sporulated T. gondii oocysts a major source of contamination for people. In the first half of 2018, an unprecedented outbreak of toxoplasmosis was reported in the city of Santa Maria, southern Brazil. The temporal and spatial distribution of the cases strongly suggested a waterborne infection. Thus, the aim of this study was to investigate a possible involvement of treated water as a source of the outbreak. For this, piglets received potentially contaminated water ad libitum for 21 days and the infection was monitored by serology through IFAT and investigation of T. gondii DNA in tissues by PCR amplification of a 529 bp followed by mouse bioassays. All piglets receiving test water ad libitum for 21 days as well as positive controls seroconverted to T. gondii. T. gondii DNA was detected in 62.5% of the piglets that received test water. All mice inoculated with tissues from each positive piglet were PCR-positive. These results strongly indicated the presence of viable oocysts in the test water administered to the animals during the study.

Combination of Adt-O1Manisa and Ad5-boIFNλ3 induces early protective immunity against foot-and-mouth disease in cattle.

Foot-and-mouth-disease (FMD) remains the most infectious livestock disease worldwide. Although commercially available inactivated or adenovirus-vectored-vaccines (Ad5-FMD) are effective, they require 5-7 days to induce protection. Therefore, new control strategies that stimulate rapid immune responses are needed. Expression of bovine interferon λ3 using the Ad5-vector platform (Ad5-boIFNλ3) is able to delay disease in cattle, but clinical signs appear at 9 days after challenge. We hypothesized that combination of Ad5-boIFNλ3 and Ad5-FMD could induce immediate and lasting protection against FMD. Cattle were vaccinated with an Ad5-FMD, Ad5-boIFNλ3, or the combination of both, followed by challenge at three days post-immunization. All animals treated with Ad5-FMD combined with Ad5-boIFNλ3 were fully protected against FMD, despite the absence of systemic neutralizing antibodies or antiviral activity at the time of challenge. Induction of a strong cell-mediated immune response suggested that Ad5-boIFNλ3 is able to act as an adjuvant of Ad5-FMD vaccine in cattle.

Evaluation of a Fiber-Modified Adenovirus Vector Vaccine against Foot-and-Mouth Disease in Cattle.

Novel vaccination approaches against foot-and-mouth disease (FMD) include the use of replication-defective human adenovirus type 5 (Ad5) vectors that contain the capsid-encoding regions of FMD virus (FMDV). Ad5 containing serotype A24 capsid sequences (Ad5.A24) has proved to be effective as a vaccine against FMD in livestock species. However, Ad5-vectored FMDV serotype O1 Campos vaccine (Ad5.O1C.2B) provides only partial protection of cattle against homologous challenge. It has been reported that a fiber-modified Ad5 vector expressing Arg-Gly-Asp (RGD) enhances transduction of antigen-presenting cells (APC) in mice. In the current study, we assessed the efficacy of a fiber-modified Ad5 (Adt.O1C.2B.RGD) in cattle. Expression of FMDV capsid proteins was superior in cultured cells infected with the RGD-modified vector. Furthermore, transgene expression of Adt.O1C.2B.RGD was enhanced in cell lines that constitutively express integrin αvß6, a known receptor for FMDV. In contrast, capsid expression in cattle-derived enriched APC populations was not enhanced by infection with this vector. Our data showed that vaccination with the two vectors yielded similar levels of protection against FMD in cattle. Although none of the vaccinated animals had detectable viremia, FMDV RNA was detected in serum samples from animals with clinical signs. Interestingly, CD4(+) and CD8(+) gamma interferon (IFN-γ)(+) cell responses were detected at significantly higher levels in animals vaccinated with Adt.O1C.2B.RGD than in animals vaccinated with Ad5.O1C.2B. Our results suggest that inclusion of an RGD motif in the fiber of Ad5-vectored FMD vaccine improves transgene delivery and cell-mediated immunity but does not significantly enhance vaccine performance in cattle.

Type III interferon protects swine against foot-and-mouth disease.

In recent years, we have developed novel strategies to control foot-and-mouth disease (FMD), including the use of biotherapeutics such as interferons (IFN) delivered by a replication-defective human adenovirus type 5 (Ad5). Swine can be sterilely protected after vaccination with an Ad5 that encodes porcine type I IFN (poIFN-α), and cattle can be similarly protected or develop significantly reduced disease when treated with an Ad5 delivering bovine type III IFN (boIFN-λ3). Here, we have evaluated the efficacy of porcine IFN-λ3 (poIFN-λ3) against FMD virus in vivo. Swine inoculated with different doses of Ad5-poIFN-λ3 were protected against disease in a dose-dependent manner. Despite the absence of systemic antiviral activity, 7 out of 10 Ad5-poIFN-λ3 inoculated animals did not develop disease or viremia, and the other 3 inoculated animals displayed delayed and milder disease by 7 days postchallenge as compared with control animals inoculated with an Ad5 control vector. While analysis of gene expression showed significant induction of IFN and IFN-stimulated genes in Ad5-poIFN-λ3-treated cultured porcine epithelial kidney cells, there was limited gene induction in peripheral blood monocytes isolated from treated swine. These results suggest that treatment with Ad5-poIFN-λ3 is an effective biotherapeutic strategy against FMD in swine.

Testes de ELISA e vírus-neutralização na detecção de anticorpos contra o vírus da diarréia viral bovina no leite / ELISA and virus- neutralization in the detection of antibodies against bovine viral diarrhea virus in milk

O sucesso na estratégia de controle e erradicação do vírus da diarréia viral bovina (BVDV), passa necessariamente pela identificação e eliminação dos animais persistentemente infectados (PI). Como esses animais excretam continuamente o vírus em suas secreções e excreções, a prevalência de anticorpos no rebanho, frequentemente é alta e com altos títulos. Devido a essas características, amostras de tanques coletivos de leite, foram submetidas a duas técnicas sorológicas, a fim de estabelecer a mais adequada na realização de triagem de rebanhos. Para isso, 767 amostras coletivas de leite foram submetidas à análise por um kit ELISA indireto (teste referência) e pela técnica de vírus-neutralização (VNT) adaptada (teste proposto). Devido aos efeitos tóxicos do leite sobre o cultivo celular, a adaptação consistiu no aumento do volume final na etapa de incubação celular. Foram positivas, 177 e 139 amostras no ELISA e na VNT, respectivamente. Com isso, a VNT adaptada apresentou uma sensibilidade de 76,8 por cento e uma especificidade de 99,5 por cento. O índice Kappa (k) foi de 0,82, demonstrando uma ótima concordância entre as duas técnicas. A análise do coeficiente de correlação entre os valores de absorbância no ELISA (OD) e os títulos de anticorpos na VNT nas amostras positivas, demonstrou uma positividade moderada (r = 0,57) com p < 0,05. No entanto, várias amostras com títulos altos na VNT apresentaram ODs moderadas ou baixas. Por outro lado, algumas amostras com títulos neutralizantes baixos apresentaram ODs altas. Como a presença de animais PI é sugerida por títulos neutralizantes ≥ 80, conclui-se que a técnica de VNT adaptada é mais adequada para a realização de triagem em amostras coletivas de leite quando objetiva-se detectar rebanhos com altos títulos de anticorpos.

The success of control/eradication programs of bovine viral diarrhea virus (BVDV) infection necessarily includes the identification and elimination of persistently infected (PI) animals. Since these animals continuously shed virus in secretions and excretions, the prevalence of antibodies in herds with PI animals is often high, and with high titers. Because of these characteristics, bulk milk samples were subjected to two serological techniques in order to establish the most appropriate in conducting screening of herds. For this, 767 bulk milk samples were analyzed by a indirect ELISA kit (reference test) and by an adapted virus neutralization (VNT) assay (proposed test). The toxic effects of milk on cell culture were reduced by increasing the final volume. One hundred seventy seven and 139 samples were positive in ELISA and VNT, respectively. Thus, the adapted VNT had a sensitivity of 76.8 percent and a specificity of 99.5 percent. The Kappa index (k) was 0.82, demonstrating an excellent agreement between the two techniques. The analysis of the coefficient of correlation between the absorbance values (OD) and VNT titers demonstrated a moderate positivity (r = 0.57). However, a significant part of samples with VNT titers ≥ 80 did not show high OD values. On the other hand, some samples with low VNT titers presented high ODs. VNT titers ≥ 80 are suggestive of the presence of PI animals in the herd. Therefore we conclude that the adapted VNT is more appropriate for herd screening when searching for herds with high antibody titers.