RhoB promotes Salmonella survival by regulating autophagy.

Salmonella enterica serovar Typhimurium manipulates cellular Rho GTPases for host cell invasion by effector protein translocation via the Type III Secretion System (T3SS). The two Guanine nucleotide exchange (GEF) mimicking factors SopE and -E2 and the inositol phosphate phosphatase (PiPase) SopB activate the Rho GTPases Rac1, Cdc42 and RhoA, thereby mediating bacterial invasion. S. Typhimurium lacking these three effector proteins are largely invasion-defective. Type III secretion is crucial for both early and later phases of the intracellular life of S. Typhimurium. Here we investigated whether and how the small GTPase RhoB, known to localize on endomembrane vesicles and at the invasion site of S. Typhimurium, contributes to bacterial invasion and to subsequent steps relevant for S. Typhimurium lifestyle. We show that RhoB is significantly upregulated within hours of Salmonella infection. This effect depends on the presence of the bacterial effector SopB, but does not require its phosphatase activity. Our data reveal that SopB and RhoB bind to each other, and that RhoB localizes on early phagosomes of intracellular S. Typhimurium. Whereas both SopB and RhoB promote intracellular survival of Salmonella, RhoB is specifically required for Salmonella-induced upregulation of autophagy. Finally, in the absence of RhoB, vacuolar escape and cytosolic hyper-replication of S. Typhimurium is diminished. Our findings thus uncover a role for RhoB in Salmonella-induced autophagy, which supports intracellular survival of the bacterium and is promoted through a positive feedback loop by the Salmonella effector SopB.

Physiological Conditions Leading to Maternal Subclinical Ketosis in Holstein Dairy Cows Can Impair the Offspring's Postnatal Growth and Gut Microbiome Development.

Maternal metabolic disruptions, such as ketosis, can have adverse effects on fetal development and influence postnatal factors. Twelve Holstein calves were randomly enrolled in this study at birth and monitored until 8 weeks of age. The study was conducted from fall 2018 until spring 2019. After completing the data collection period, calves were classified according to their respective dams ketotic condition after parturition. This classification was based on dam blood ß-hydroxybutyrate < 1.4 mmol/L nonketotic (NONKET; n = 6 calves) or ≥1.4 mmol/L subclinical-ketotic (SK; n = 6 calves). SK calves had greater birth body weight (p = 0.05) but exhibited a slower growth rate compared to NONKET calves from 1 to 8 weeks (p = 0.02). At birth, SK calves had lower (p < 0.01) levels of non-esterified fatty acids and bilirubin compared to NONKET calves. Analysis of feces alpha diversity indicates that by 3 weeks, NONKET calves had greater diversity, richness, and evenness. Butyricicoccus pullicaecorum and Gallibacterium anatis were more abundant in SK calves (p < 0.05) at 3 weeks. In contrast, NONKET calves had a greater (p < 0.05) abundance of Sharpae azabuensis at 3 weeks. These findings suggest that subclinical ketosis in cows can impact the in-utero development, postnatal growth, and maturing gut microbiome of their offspring.

Yeast Culture Supplementation Effects on Systemic and Polymorphonuclear Leukocytes' mRNA Biomarkers of Inflammation and Liver Function in Peripartal Dairy Cows.

This study evaluated the effects of feeding a commercial yeast culture on blood biomarkers and polymorphonuclear leukocyte (PMNL) gene expression in dairy cows during the transition period until 50 d postpartum. Forty Holstein dairy cows were used in a randomized complete block design from -30 to 50 d. At -30 d, cows were assigned to a basal diet plus 114 g/d of top-dressed ground corn (control; n = 20) or 100 g/d of ground corn and 14 g/d of a yeast culture product (YC; n = 20). Blood samples were collected at various time points from -30 to 30 DIM to evaluate blood biomarkers and PMNL gene expression related to inflammation, liver function, and immune response. Liver function biomarkers, gamma-glutamyl transferase (GGT) and albumin were greater and lower, respectively, in YC cows in comparison to control. However, these biomarkers remained within physiological levels, indicating an active inflammatory process. Genes in PMNL expression related to inflammation (NFKB1, TNFA, TRAF6), anti-inflammation (IL10), and cell membrane receptors (SELL) were upregulated in the YC group in comparison to control. These results suggest that YC could stimulate a more active inflammatory response with signs of a resolution of inflammation in transition cows.

Relationship between thermal comfort indices and internal temperature of grazing lactating Holstein × Gyr cows in western Amazonia / Relação entre índices de estresse térmico e a temperatura interna de vacas Holandês x Gir lactantes em pastejo na Amazônia Ocidenta

The relationship between the indices of thermal comfort and internal temperature in lactating cows was evaluated inRondônia, western Amazonia in a 2 × 2 crossover trial carried out with eight Girolando cows, ½ Holstein × ½ Gyr (n=4) and Holstein × » Gyr (n=4), grazing palisade grass supplemented with or without soybean oil concentrates. Data logger thermometers adapted to intravaginal devices (hormone free) were used to measure internal temperatures (IT) every 10 min over 48 h. Concomitantly, environmental parameters (air temperature, relative humidity, wind speed, and solar radiation) were collected for the estimation of the temperature-humidity index (THI), equivalent temperature index (ETI), and predicted respiratory rate (PRR). Supplementation with soybean oil did not affect the internal temperature of the cows. IT varied significantly between the genetic groups, with the highest mean observed at night in the Holstein × » Gyr cows, compared with that in the ½ Holstein × ½ Gyr cows (39.54 °C vs 39.06 °C, respectively). THI, ETI, and PRR were significantly and positively correlated with the IT of the Girolando cows; however, the PRR was not found to be adequate in indicating the night-time heat stress in cows.

A relação entre índices de conforto térmico com a temperatura interna de vacas lactantes foi avaliada em Rondônia, na Amazônia Ocidental, em um ensaio crossover 2x2 utilizando oito vacas Girolando, ½ Holandês x ½ Gir (n=4) e Holandês x » Gir (n=4) pastejando capim-marandu suplementadas com concentrados contendo ou não óleo de soja. A temperatura interna (TI) dos animais foi aferida com termômetros eletrônicos adaptados em dispositivos intravaginais (livres de hormônio) a cada 10 minutos durante 48h. Concomitantemente, dados climáticos (temperatura do ar, umidade relativa do ar, velocidade do vento e radiação solar) foram coletados para cálculo dos índices de temperatura e umidade (ITU), índice de temperatura equivalente (ITE) e o índice de frequência respiratória predita (PFR). A suplementação com óleo de soja não afetou a temperatura interna das vacas. A TI variou significativamente entre os grupos genéticos, com a maior média observada à noite nas vacas Holandês × » Gir em relação às ½ Holandês × ½ Gir (39,54 °C vs 39,06 °C, respectivamente). ITU, ITE e PFR tiveram correlação significativa e positiva com a TI de vacas Girolando, mas o PFR não foi adequado para indicar estresse térmico em vacas no período da noite.

Characterization of single nucleotide polymorphisms in the promoter region of the bovine paraoxonase 1 (PON1) gene affecting serum enzyme activity in dairy cows.

The aim of the present study was to characterize single nucleotide polymorphisms (SNPs) in the promoter region of the bovine paraoxonase 1 (PON1) gene and determine their relationship with PON1 serum activity. Blood samples were collected from 47 Holstein cows during the periparturient period. Serum PON1 was measured and DNA extracted for PCR and sequencing of the promoter region of the PON1 gene. Seven novel SNPs were identified and of these, five SNPs, at positions -105, -221, -392, -611 and -674, upstream of the start of the published mRNA sequence (NM_001046269.2), were found to be associated with variability in serum PON1 activity (P < 0.05). The SNPs at positions -221 and -611 were located in regions predicted to bind to transcription factors linked to the acute phase response.