Genetic background of CTX‐M‐15‐producing Enterobacter hormaechei ST114 and Citrobacter freundii ST265 co‐infecting a free‐living green turtle (Chelonia mydas)

CTX‐M‐type extended‐spectrum β‐lactamase (ESBL)‐producing Enterobacteriaceae have become identified in marine ecosystem constituting a serious ecological issue. In this respect, although contamination of coastal waters and seafood, and even colonization of seabirds and fishes have been increasingly reported, molecular data are lacking to elucidate the clinical impact of ESBL producers in infected marine animals. In this study, using a genomic approach, we have analysed the genetic background of CTX‐M‐15‐producing Enterobacter hormaechei (belonging to the international human clone ST114) and Citrobacter freundii (ST265) co‐infecting a free‐living green turtle (Chelonia mydas) suffering from septic arthritis, which progressed to generalized coelomitis and death. Wide resistome of these pathogens contributed to treatment failure and death of the animal.     

Nitric oxide impacts bovine sperm capacitation in a cGMP‐dependent and cGMP‐independent manner

We aimed to elucidate whether NO acts in in vitro sperm capacitation in bovine via cGMP/PKG1 pathway. For this, cryopreserved bovine sperm were capacitated in vitro with 20 µg/ml heparin (Control) plus treatments: 1 mM L‐arginine (L‐arg, NO precursor), 50 µM Rp‐8‐Bromo‐β‐phenyl‐1,N²‐ethenoguanosine‐3′,5′‐cyclic monophosphorothioate (Rp‐8‐Br‐cGMPS, selective inhibitor of the binding site for cGMP in PKG1), 1 mM 2‐Phenyl‐4,4,5,5‐tetramethylimidazoline‐1‐oxyl 3‐oxide (PTIO, NO scavenger), and the combinations of L‐arg + RP‐8‐Br‐cGMPS and L‐arg + PTIO. Sperm motility and vigour were determined by phase‐contrast microscopy, capacitation status by chlortetracycline staining, and the intracellular concentration of cGMP was measured by ELISA. Data were subjected to analysis of variance and means compared with SNK test at 5% probability. Motility and vigour were lower in sperm treated with PTIO when compared to Control and other treatments (p < .05). The L‐arg treatment showed the highest percentage of capacitated sperm when compared to the Control and other treatments (Rp‐8‐Br‐cGMPS, L‐arg + Rp‐8‐Br‐cGMPS and PTIO) (69.8 ± 3.4%, 51.2 ± 3.0, 51.1 ± 2.1, 51.2 ± 3.0 and 45.5 ± 2.7, respectively) (p < .05). The capacitation ratio (%) was lower in treatments with Rp‐8‐Br‐cGMPS, L‐arg + Rp‐8‐Br‐cGMPS and PTIO, respectively (p < .05). Lastly, cGMP concentration (pmol/ml) was lower in PTIO and L‐arg + PTIO (1.3 ± 0.3 and 1.6 ± 0.4) and was higher in Rp‐8‐Br‐cGMPS and L‐arg + Rp‐8‐Br‐cGMPS (3.7 ± 0.4 and 4.0 ± 0.5) treatments. We showed that during in vitro capacitation of cattle: (a) NO influences sperm motility and vigour; (b) NO is associated with cGMP synthesis through two independent pathways and (c) the cGMP/PKG1 pathway has a partial role in sperm capacitation and does not involve the L‐arg/NO.     

Nutritional evaluation and productivity of supplemented sheep grazing in semiarid rangeland of northeastern Brazil

Tropical Animal Health and Production - Sheep production systems in Brazilian caatinga rangelands require supplementation adapted to changes in floristic and chemical composition as dry seasons...     

Genetic diversity,anti‐microbial resistance,plasmid profile and frequency of the Vi antigen in Salmonella Dublin strains isolated in Brazil

Salmonella Dublin is strongly adapted to cattle causing enteritis and/or systemic disease with high rates of mortality. However, it can be sporadically isolated from humans, usually causing serious disease, especially in patients with underlying chronic diseases. The aim of this study was to molecularly type S. Dublin strains isolated from humans and animals in Brazil to verify the diversity of these strains as well as to ascertain possible differences between strains isolated from humans and animals. Moreover, the presence of the capsular antigen Vi and the plasmid profile was characterized in addition to the anti‐microbial resistance against 15 drugs. For this reason, 113 S. Dublin strains isolated between 1983 and 2016 from humans (83) and animals (30) in Brazil were typed by PFGE and MLVA. The presence of the capsular antigen Vi was verified by PCR, and the phenotypic expression of the capsular antigen was determined serologically. Also, a plasmid analysis for each strain was carried out. The strains studied were divided into 35 different PFGE types and 89 MLVA‐types with a similarity of ≥80% and ≥17.5%, respectively. The plasmid sizes found ranged from 2 to >150 kb and none of the strains studied presented the capsular antigen Vi. Resistance or intermediate resistance was found in 23 strains (20.3%) that were resistant to ampicillin, ciprofloxacin, chloramphenicol, imipenem, nalidixic acid, piperacillin, streptomycin and/or tetracycline. The majority of the S. Dublin strains studied and isolated over a 33‐year period may descend from a common subtype that has been contaminating humans and animals in Brazil and able to cause invasive disease even in the absence of the capsular antigen. The higher diversity of resistance phenotypes in human isolates, as compared with animal strains, may be a reflection of the different anti‐microbial treatments used to control S. Dublin infections in humans in Brazil.     

Genetic‐Quantitative Study of the First‐Service Pregnancy Probability of Murrah Heifers

Because of the importance of reproduction in stock breeding systems, it is necessary to find selection criteria that increase reproductive efficiency. The aim of this study was to estimate genetic parameters for the probability of conception on first service (PROB) in Murrah heifers, and its association with other traits of economic interest [age at first calving (AFC), service period, calving interval and milk yield at 270 days], with the purpose of evaluating their use as selection criteria. Reproductive information and first lactation records of 1200 Murrah heifers were used to perform two‐trait analyses between PROB and the other characteristics. Bayesian inference was used to estimate the variance components, considering PROB as threshold and the other as linear factors. The results demonstrate that this trait has heritability of 0.15, indicating the possibility of a genetic gain by using it for selection. With respect to the genetic correlation estimates, the only high‐magnitude association was with AFC (?0.899), which is the current criterion indicating sexual precocity of females. In the light of the parameters estimated, the first‐service pregnancy rate is an alternative for indication of sexual precocity, although presenting a smaller genetic gain than the current standard AFC. Nevertheless, additional research should be conducted regarding this trait to assess the economic importance of its use in dairy buffalo production systems.     

Sperm cryopreservation of tiete tetra Brycon insignis (Characiformes): effects of cryoprotectants,extenders, thawing temperatures and activating agents on motility features

The aim of this study was to test the effects of cryoprotectants [dimethyl sulphoxide (DMSO) and methylglycol], extenders (0.9% NaCl, 5% glucose, Beltsville Thawing Solution^? and Merck III^?), thawing temperatures (30 and 60 °C) and activating agents (0.29% NaCl and 1% NaHCO₃) on the cryopreservation process of tiete tetra Brycon insignis sperm. Sperm was loaded in 0.5 mL straws, frozen in nitrogen vapour at ?170 °C and stored in liquid nitrogen. Post‐thaw sperm quality was evaluated in terms of subjective motility rate, quality motility score (0=no movement; 5=rapidly swimming spermatozoa), duration of motility and vitality (eosin–nigrosin staining). Post‐thaw sperm motility rate was greater in methylglycol (76–88%), compared with DMSO (23–59%). In general, the highest quality motility scores were observed when sperm was thawed at 30 °C and triggered in 1% NaHCO₃ (3.5–4.3). Duration of motility was longer when triggered in 1% NaHCO₃ (95–120 s) compared with 0.29% NaCl (69–107 s). Sperm vitality was not affected by any of the parameters tested and varied from 51% to 69% intact sperm. Brycon insignis sperm frozen in methylglycol combined with any of the extenders tested and using the methods described above yields motility above 57% and that should last long enough to fertilize oocytes.     

Liver histology and histomorphometry in hybrid sorubim (Pseudoplatystoma reticulatum × Pseudoplatystoma corruscans) reared on intensive fish farming

This study aimed to characterize the liver histology and histomorphometry in sorubim hybrid of different categories (nursery, growth and grow‐out) reared on fish farming. The categories were defined considering body weight (BW): nursery category (n = 5): BW = 37.06 ± 6.00 g (31.6–45.3 g); growth category (n = 5): BW = 310.40 ± 53.80 g (242.1–376.4 g) and grow‐out category (n = 5): BW = 874.28 ± 27.59 g (846.2–913.1 g). Liver fragments were processed to paraffin inclusion, and sections were stained by haematoxylin and eosin (H&E), PAS (Periodic Acid Schiff) and Perl's staining to histology, histomorphometry and density volumetric of liver structures; glycogen analysis and to detect ferric irons (Fe³⁺) respectively. The hepatosomatic index decreased between the categories (P < 0.01). The percentage of PAS‐positive hepatocytes in the nursery category was higher (P < 0.05) in relation to the growth and grow‐out categories. The hepatocytes from all fish were positive to Perl's staining. The density volumetric of liver structures did not differ among categories except to blood vessels were higher (P < 0.01) in the nursery and growth. The area (μm²) and perimeter (μm) of hepatocytes, and the area (μm²), perimeter (μm) and volume (μm³) of the nuclei from grow‐out fish were lower (P < 0.01) than those from the nursery and growth categories. Changes in morphometric characteristics of hepatocytes may result from metabolic changes associated with body growth surubins; therefore, these morphometric characteristics of liver tissue can be used as functional biomarkers for the assessment of fish health and nutrition status.     

Aurantiochytrium sp. meal as DHA source in Nile tilapia diet,part II: Body fatty acid retention and muscle fatty acid profile

Nile tilapia juveniles (8.35 ± 0.80 g) were fed on four levels (0.0%; 0.5%; 1.0%; 2.0%, 4.0%) of Aurantiochytrium sp. meal (ALL‐G‐RICH?), a source of docosahexaenoic acid (DHA). The 1% Aurantiochytrium sp. meal diet was compared to a control diet, which contained the same amount of DHA as cod liver oil (CLO) at 1.7% diet. Groups of 25 fish were stocked in 100 L tanks and fed twice daily until apparent satiation, for 57 days, at 28°C. Increasing dietary Aurantiochytrium sp. meal reduced the body retention of DHA and n‐3 polyunsaturated fatty acids (n‐3 PUFA) but increased the body retention of alpha‐linolenic (α‐LNA), linoleic (LOA) and n‐6 polyunsaturated fatty acids (n‐6 PUFA). Fatty acid profile in tilapia muscle was affected by increasing dietary inclusions of Aurantiochytrium sp. meal, with an increase in DHA, α‐LNA, n‐3 PUFA and n‐3 long chain‐polyunsaturated fatty acids (n‐3 LC‐PUFA) but a decrease in monounsaturated fatty acids (MUFA), n‐6 PUFA and n‐6 long‐chain polyunsaturated fatty acids (n‐6 LC‐PUFA). There was a larger body retention of DHA, α‐LNA, LOA, n‐3 PUFA and n‐6 PUFA fatty acids and a higher percentage of DHA, n‐3 PUFA and n‐3 LC‐PUFA in muscle fatty acid profile in fish fed on CLO diets than in those fed on 1% Aurantiochytrium sp. Therefore, Aurantiochytrium sp. meal is an alternative source of DHA for Nile tilapia diets.     

Dietary organic acids blend alone or in combination with an essential oil on the survival,growth, gut/liver structure and de hemato‐immunological in Nile tilapia Oreochromis niloticus

This study evaluated the effects of dietary supplementation of organic acids blend (OAB) alone or in combination with essential oil, Lippia origanoides (OAE) for Nile tilapia fed supplemented diets for 30 days. Fish (1.1 ± 0.04 g) were fed control (Control), or OAB 0.5% or OAB 0.5% + essential oil 0.125% (OAE) respectively. At the end of the experiment, samples were collected for de hemato‐immunological, histological analysis of the intestine and liver, as well as microbiology of the intestine. The pH of the diets supplemented with OAB and OAE reduced 0.92 and 0.19 respectively. The growth and FCR were unaffected by the treatments, but survival was significantly higher in the OAB treatment. Fish fed the OAB diet showed reduced concentration of total heterotrophic bacteria and Pseudomonas sp. in the intestine. Increased glucose in fish fed OAB and high number of circulating monocytes in fish fed OAE diet were observed. The anterior intestine of fish fed OAE diet showed larger number of goblet cells and increased villi height. The diet supplemented with OAB, mainly, improved the intestinal health and survival of tilapia juveniles and can be used in juvenile production.