Bacterial community associated with the ambrosia beetle Platypus cylindrus on declining Quercus suber trees in the Alentejo region of Portugal

In Portugal, the oak pinhole borer Platypus cylindrus and its mycobiota have been associated with cork oak (Quercus suber) death, but no knowledge exists regarding the associated bacterial community. However, it is known that some bacteria are important for ambrosia beetle symbiosis and play a role in oak tree health. To explore the bacteria associated with this beetle and its host, with the ultimate goal of highlighting potential roles in oak decline, this study used a culture-dependent approach for strain isolation and phylogenetic identification using 16S rRNA gene sequencing and multilocus sequence analysis (MLSA). The bored galleries of different cork oak trees from a cork stand in Alentejo, together with the body and mycangia of adult beetles, were investigated. The samples revealed a diverse community comprising 500 isolates with 64 distinct types of bacterial colonies. Sixty-eight strains were selected for sequencing and used for phylogenetic analysis, 40 from wood galleries and 28 from beetles. Thirty-two genera of bacteria were identified, 18 of which were described for the first time within oak–beetle interactions. Major taxonomic groups were Actinobacteria in beetles and Enterobacterales in wood galleries. Although specific oak bacterial pathogens were not detected, a group of distinct strains detected in wood galleries, potentially belonging to a new Pectobacteriaceae species, were able to produce mild symptoms on cork oak plantlets. This study reports for the first time the biodiversity of culturable bacteria associated with the Q. suber–P. cylindrus interaction, their relevance to both organisms and the possible contribution to oak decline.     

Biosorption of macronutrients by Brazilian tropical peats

The objective of this work was to evaluate the adsorption of macronutrients calcium, potassium, magnesium, nitrogen, and phosphorus in two Brazilian tropical peat samples, investigating the effect of pH and determining the kinetics of the adsorption process. Two different Brazilian tropical peat samples were characterized using FTIR, TG, and SEM techniques. Different pH conditions were tested, as well as different mass concentrations of the peats. Differences in the chemical structures of the peat samples directly influenced the adsorptive capacities for the macronutrients. The adsorptive capacity for nitrogen was highest at pH 3, while the best adsorption of calcium and potassium was obtained at pH 6. The best fit to the data was provided by the pseudo-second-order model, which confirmed the rapid adsorption of calcium by both peats.     

Presence of Leptospiral DNA in Semen Suggests Venereal Transmission in Horses

The purpose of the present study was to detect leptospiral DNA by PCR in semen and urine samples of stallions to test for venereal transmission in horses. A total of 10 stallions from four herds were studied, and sampling was conducted in semen and urine for culture and PCR and serum for serology. From the 10 serum samples tested, 6 (60%) were seroreactive. No pure culture was obtained, but leptospiral DNA was detected by PCR in 50% of the semen samples and 30% of urine samples. The present study aimed to detect leptospiral DNA by PCR in semen and urine samples of stallions to test for venereal transmission in horses. Based on these findings, we suggest that there is potential transmission of leptospirosis in horses by sexual transmission.     

In vitro development of IVF-derived bovine embryos following cytoplasmic microinjection for the episomal expression of the IGF2 gene

Important genomic imprinting changes usually occur following the in vitro production (IVP) of bovine embryos, especially in the imprinting pattern of components of the IGF system. This study aimed to evaluate the effects of a transient episomal overexpression of the IGF2 gene in bovine IVP embryos following embryo cytoplasmic microinjection (CMI) at the 1-cell stage on embryo survival, early and late developmental kinetics and morphological quality up to Day 7 of development. Selected cumulus–oocyte complexes (COCs) were matured and fertilized in vitro and subsequently segregated into six experimental groups: non-CMI control group and five CMI groups at increasing doses (0, 10, 20, 40 and 80 ng/μl) of a GFP vector built for the episomal expression of bovine IGF2. Zygote CMI was effective in delivering the expression vector into the ooplasm, irrespective of the groups, with 58% of positive GFP fluorescence in Day 7 blastocysts. Considering developmental rates and late embryo kinetics, the 10-ng/μl CMI vector dose promoted a lower blastocyst rate (10.4%), but for blastocysts at more advanced stages of development (93.0% blastocysts and expanded blastocysts), and higher number of cells (116.0 ± 3.0) than non-CMI controls (23.3%, 75.0% and 75.0 ± 6.8 were obtained, respectively). In conclusion, CMI at the 1-cell stage did not compromise subsequent in vitro development of surviving embryos, with the 10-ng/μl group demonstrating a possible growth-promoting effect of the IGF2 gene on embryo development, from the 1-cell to the blastocyst stage.     

Evaluation of the Effective Inactivation of Enteric Bacteria and Viruses From Swine Effluent and Sludge at Tropical Temperatures

The safe recycling of organic wastes such as the nutrients obtained from agriculture activities is a relevant aspect of the “One Health” strategy, a worldwide initiative including all aspects of health care for humans and animals, as well as aspects related to the conservation and development of the environment. It represents an important opportunity to mitigate the potential impact of microbial pathogens likely present in nutrient natural sources, as digestate, when applied to land as fertilizers. The objectives of the current study were to determine the inactivation of enteric microorganisms in swine digestate (effluent and sludge) during storage in anaerobic conditions and at different ambient temperatures of tropical zones (16, 22, and 37 °C). Human adenovirus 2 (HAdV-2), bacteriophage PhiX-174, and Salmonella enterica serovar typhimurium (S. typhimurium) were used as enteric microbial models to determine the minimum storage time required to reduce at least 3log of enteric microorganisms load in both matrices at the mentioned temperatures. The minimum storage time required for S. typhimurium was shorter than those observed for viruses at 37 °C, while reduction of at least 3log for S. typhimurium was observed after just 1 day; 20 and 90 days were needed for similar reductions for HAdV-2 and PhiX-174, respectively. Similar results were also observed at lower temperatures: 30 and 45 days were needed for S. typhimurium at 22 and 16 °C, respectively, but 30 and 90 days were needed at 22 °C for HAdV-2 PhiX-174, respectively, or 90 and 120 days at 16 °C for HAdV-2 PhiX-174, respectively. This is a pioneering study on the evaluation of inactivation of enteric viruses in swine digestate in usual tropical ambient temperatures; it demonstrates that the time for a considerable inactivation is longer than that observed for a enteric microorganisms (S. typhimurium), and shows that the storage of swine digestate at the referred temperatures can be used as a low-cost post-treatment to inactivate enteric microorganisms, allowing a further use as safer fertilizer.     

Genotyping of Mycobacterium bovis by geographic location within Mexico

The spacer oligonucleotide typing (spoligotyping) method was used to differentiate 62 Mycobacterium bovis isolates obtained from tissues with macroscopic lesions typical of tuberculosis in dairy cattle from different regions of Mexico. Our purpose was to see if a strain from one region was genetically different from those of other regions (with the long-term aim of doing molecular trace back of isolates obtained in the laboratory). Results from the genetic analysis indicate that M. bovis isolates cannot be grouped by geographic location due to a wide range of genetic types involved in dairy cattle infections. Isolates even from the same herd showed different spoligotypes but some isolates from different region had similar genetic patterns. Genetic typing without epidemiologic information does not seem to be a plausible method to trace back animals to source of origin to detect and eliminate sources of infection.     

Carcass traits and meat quality of Texel lambs raised in Brachiaria pasture and feedlot systems

Our hypothesis suggests an improvement in carcass and meat quality of pasture‐finished animals by introducing concentrate feed into diets. This study aimed to evaluate the effect of different levels of protein‐energy supplementation on carcass and meat characteristics of Texel lambs raised on Brachiaria pastures, and compare the results with those obtained from confined animals. Thirty 2‐month‐old intact lambs were divided into five treatments: pasture with mineral supplementation, pasture containing daily supplies of 0.8, 1.6, and 2.4% BW protein‐energy supplementation, and confinement (reference treatment). All animals were slaughtered 104 days after the beginning of the experiment. Lambs submitted to 1.6% and 2.4% BW supplementation presented similar characteristics to animals kept in confinement and were superior to animals treated with 0 or 0.8% of BW supplementation levels (p < .05). Increases of subcutaneous fat thickness were verified for 0.0, 0.8, 1.6, 2.4% BW, and confinement treatments, being respectively of 0.25, 0.74, 1.61, 1.69, and 1.98. Conversely, treatments had no influence on meat physical‐chemical characteristics, being all considered moderately soft, juicy (tender), and with mild tastes and odors. Lambs raised on Brachiaria pastures have high‐quality meat, but more protein‐energy supplementation is needed to be an alternative for the feedlots for young animals.     

Influence of the duration of calving and obstetric assistance on the placental retention index in Holstein dairy cows

The aim of this study is to evaluate the influence of duration of calving and obstetric assistance on retained placenta incidence of high milk production Holstein cows. Experimental groups were determined according to the duration of the expulsive phase of calving: 2 h (n = 16), 2‐4 h (n = 16) and >4 h (n = 12), and additionally allocated in two sub‐groups: spontaneous calving (n = 22) and intervention calving (n = 22). Diagnosis of retained placenta was considered with a threshold of 8 h after the expulsive phase. Cows without obstetric intervention, with labor duration of 2 h, presented reduced time needed for placental release, in comparison to those with obstetric assistance. In the 2–4 h group and >4 h, there was no statistical difference. The 2 h and 2–4 h groups with intervention and the spontaneous >4 h group were considered retained placenta groups. On the other hand, performing obstetric intervention when calving period was superior to 4 h nulled the occurrence of retention of fetal membranes. As a conclusion, obstetric assistance predisposes placental retention to calving with <2 h of duration in dairy cows. Conversely, when calving is more than 4 h, performing fetal extraction has a beneficial influence on preventing retained placenta.