Streptococcus suis in employees and the environment of swine slaughterhouses in S?o Paulo,Brazil: Occurrence,risk factors,serotype distribution,and antimicrobial susceptibility

Streptococcus suis is an important pathogen in the swine industry. This article is the first to report the occurrence, risk factors, serotype distribution, and antimicrobial susceptibility of S. suis recovered from employees and environmental samples of swine slaughterhouses in Brazil. Tonsillar swabs from all 139 pig-slaughtering employees and 261 environmental swabs were collected for detection of S. suis and serotyping by monoplex and multiplex polymerase chain reaction, respectively. Antimicrobial susceptibility was determined by the disk-diffusion method. Although S. suis was not detected in any of the tested employees, it was isolated from 25% of the environmental samples. Significant differences (P < 0.05) in the occurrence of S. suis were observed between slaughterhouses and between areas of low, medium, and high risk. The most frequent serotypes were 4 and 29, each accounting for 12% of the isolates, followed by 5, 12, 21, and 31, each accounting for 6%. High rates of susceptibility to the antimicrobials doxycycline (100%), ceftiofur (94%), ampicillin (81%), and cephalexin (75%) were observed. However, multidrug resistance was observed in all the isolates. Because S. suis is present in the environment of swine slaughterhouses, on carcasses and knives, as well as on the hands of employees in all areas, all employees are at risk of infection.     

The Effect of Oxidative Stress on Thawed Bulk‐Sorted Red Deer Sperm

The aims of this study were to assess the effects of the sex‐sorting process on post‐thaw sperm quality as well as on induced oxidative stress damage (H₂O₂ 0 mm  = H000; H₂O₂ 50 mm  = H050; H₂O₂ 100 mm  = H100) and the protective action of reduced glutathione (GSH) and Trolox, when comparing sorted (BSS) and non‐sorted (NS) red deer spermatozoa incubated at 37°C. Sperm samples from three stags were collected by electroejaculation and frozen. Immediately after thawing, sperm motility was higher (p < 0.05) for NS (59% ± 3.3) than BSS (36.9% ± 5.8) sperm. Furthermore, the percentage of apoptotic sperm was higher (p < 0.05) for BSS (21.6% ± 5.0) than NS sperm (14.6% ± 1.2). The presence of H₂O₂ increased DNA damage in NS (H000 = 4.1% ± 0.9; H050 = 9.3% ± 0.7; and H100 = 10.9% ± 2.3), but not in BSS sperm. However, in the presence of oxidant, GSH addition improved (p < 0.05) sperm motility in both groups of sperm samples as compared to their controls (NS: 44.5 ± 4.8 vs 21.1 ± 3.9 and BSS: 33.3 ± 8.1 vs 8.9 ± 1.8). These results demonstrate that the sperm‐sorting process induces sublethal effects, albeit selecting a sperm population with a chromatin more resistant to oxidative stress than that in non‐sorted sperm. Moreover, addition of GSH at 1 mm may be a good choice for maintaining the quality of stressed sperm samples, unlike Trolox, which inhibited sperm motility.     

Lactoferrin increases sperm membrane functionality of frozen equine semen

During cryopreservation, sperm was submitted to an increase in reactive oxygen species generation. This work aimed to improve the quality of frozen equine sperm after the addition of antioxidants lactoferrin (Lf) and catalase (Cat) to a freezing extender. Semen from six stallions was frozen with the extenders: F1) control, INRA 82 freezing extender, F2) F1 + 500 μg/ml Lf and F3) F1 + 200 IU/ml Cat. After thawing, sperm motility parameters, membrane functionality and integrity, and acrosome integrity and spontaneous acrosome‐reacted sperm were evaluated with a computer‐assisted sperm analysis, a hypoosmotic swelling test and epifluorescent microscopy, respectively. Nitrite, hydroperoxide and iron concentrations of frozen semen were measured with spectrophotometry. The percentage of functional membrane sperm treated with Lf was higher (50.7% ± 11.6%) compared to that of the control (37.6% ± 15.6%), while the iron (61.4 ± 11.6 vs 73.3 ± 13.8 mg/dl) and nitrite concentrations (16.3 ± 7.1 vs 25.9 ± 4.2 μM/μg protein) were lower, respectively (p < .05). Thus, it can be suggested that Lf protect stallion spermatozoon during freezing as it has increased the percentage of sperm with functional membrane and decreased the lipid oxidant agents.     

Human disturbance affects enzyme activity,microbial biomass and organic carbon in tropical dry sub-humid pasture and forest soils

ABSTRACT

The Brazilian dry forest occupies an area of about 1 million km² approximately 46% of which has been deforested. Many studies have been done on the effects of this on productivity and soil chemical attributes. However, little is known about soil enzymatic activity, which is sensitive to environmental changes. The objective of this study was to evaluate the effects of deforestation using different levels of human disturbance on soil enzyme activities, organic carbon content, microbial biomass, and microbial community. We studied areas covered with forest (TDF), old grass (OG), or new grass (NG). Soils from NG had increased microorganisms, which restored important processes related to carbon, sulfur, and nitrogen cycling, so that they resembled those in the forested area. The results of this study showed that the conversion of forest to pasture with a high level of human disturbance could decrease the activities of β-glucosidase, urease, alkaline phosphatase, and fluorescein diacetate in the soil by up to 87%, 66%, 62%, and 58%, respectively. These findings suggest that human disturbance can cause substantial changes in the enzymatic activity and microbial community in the soil. We suggest that maintaining grass pasture with low human disturbance should have fewer impacts on soil quality.     

Toxoplasma gondii diagnosis in ovine aborted fetuses and stillborns in the State of Pernambuco, Brazil

The aim of this research was to study the contribution of Toxoplasma gondii to reproductive failure using nested PCR and histopathological examination of fetuses, stillborns and placentas. We examined 245 organs of fetuses and 28 placentas from 35 abortions and stillborns from naturally occurring miscarriages in sheep in the State of Pernambuco, Brazil. At necropsy, fragments of brain, cerebellum, medulla, lung, heart, spleen, liver and placenta were taken for nested PCR and histopathological tests. Pathological examination revealed macroscopic lesions, suggesting T. gondii infection in 5/35 (14.3%) of the placentas. The histopathological examination revealed no lesions characteristic of toxoplasmosis in the organs investigated. In the five placentas, lesions consistent with toxoplasmosis were observed as an inflammatory non-suppurative infiltrate, along with multiple necrosis and mineralization. Nested PCR showed three aborted fetuses and two stillborns (14.3%) to test positive for T. gondii, with DNA amplification in all organs and the placenta, especially the heart and the placenta, which are the tissues of choice. This study substantiates the theory that T. gondii is involved in miscarriages and stillbirths and in the placentas of naturally infected sheep in Brazil. Such findings have not previously been described in the national literature.     

A shot in the dark for conservation: Evidence of illegal commerce in endemic and threatened species of elasmobranch at a public fish market in southern Brazil

1. The illegal sale of endangered elasmobranchs has been recorded in a number of different Brazilian states, where sharks and rays are being marketed primarily as ‘cação’ or ‘viola’. Brazil is ranked among the top 10 nations worldwide that harvest most sharks, causing an immeasurable impact on the local elasmobranch populations.
2. The present study applied the DNA barcoding technique, based on the cytochrome C oxidase subunit I gene, for the molecular identification of the elasmobranch species sold as processed products under the generic names of ‘cação’, ‘mangona’, ‘azul’, ‘cambeva’, ‘fiuso’ and ‘lombo preto’, in the fish market of the city of Florianópolis, capital of the state of Santa Catarina, Brazil.
3. Nine elasmobranch species were identified in the 56 samples analysed, including six sharks and three rays, representing six families, the Carcharhinidae, Sphyrnidae, Squatinidae, Arhynchobatidae, Myliobatidae and Gymnuridae. Prionace glauca, identified in more than half (56%) of the samples analysed, is listed as Near Threatened by the International Union for the Conservation of Nature.
4. Six species identified in the present study, Sphyrna zygaena, Sphyrna lewini, Squatina guggenheim, Carcharhinus signatus, Gymnura altavela and Rioraja agassizii, are under some level of risk of extinction, while two others (Rhizoprionodon lalandii and Myliobatis goodei) are listed as Data Deficient.
5. Our results indicate that the commercial exploitation of endemic sharks and rays at risk of extinction is commonplace in southern Brazil. This reinforces the need for more systematic monitoring of the trade in fishery products and more effective application of the environmental legislation and conservation programmes.

     

Morphofunctional Characterization of Cooled Sperm With Different Extenders to Use in Equine-Assisted Reproduction

The aim of this study was to assess extenders for cooling equine semen at 5°C and to be used in assisted reproductive technologies (ARTs). Four ejaculates were obtained from each of four stallions. Gel-free semen was diluted in three different extenders: (1) SMK, an opaque skim milk–based extender; (2) SMT, a skim milk (65%) and Tyrode medium (35%); and (3) BSAG, a clear extender containing 1% bovine serum albumin. Samples were packaged (10 mL; 50 × 10⁶ sperm/mL) and stored in a cooling device at 5°C for 12 hours. Analyses were done at 0, 4, 8, and 12 hours after cooling. Semen was analyzed for sperm motility characteristics using a computer-assisted sperm analysis, for plasma membrane and acrosome integrity and mitochondrial membrane potential, using fluorescent probes (propidium iodide, Hoechst 33342, fluorescein isothiocyanate–conjugated Pisum sativum agglutinin, and 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolyl carbocyanine iodide [JC-1]). Morphology was evaluated with differential interference contrast microscopy and sperm chromatin integrity by the toluidine blue technique. Data were analyzed by analysis of variance and by Tukey test, with time as a repeated measure (SAS, 1998), when P < .05 was significant. In general, milk-based extenders (SMT and SMK) showed improved maintenance of semen quality compared with BSAG. Finally, the addition of skim milk to equine semen extender for cooling at 5°C for 12 hours seems to play a crucial role in sperm preservation. Although, optically clearer extenders are desired for use in ARTs, such as sperm sexing, the milk-free extender (BSAG) is less efficient for cooled-stored equine semen.     

Melatonin affects conjugation of 4-hydroxynonenal with glutathione in liver of pacu, a hypoxia-tolerant fish

In cytosol from liver of pacu, Piaractus mesopotamicus, a hypoxia-tolerant fish that dwells in Pantanal, we found an enzyme activity capable of modulating the alkenal 4-hydroxy-2-nonenal (HNE) by conjugating it with glutathione (GST–HNE activity). HNE is a downstream metabolite from the oxidation of polyunsaturated fatty acids by reactive oxygen species arisen from mitochondria of animal cells. HNE production may increase more intensively under oxidative stress. Harmful effects to cell survival may occur when HNE increases over 10^?4 M. Pacus submitted to hypoxia in July (cold season in Pantanal) showed 40 % less of this GST–HNE conjugating activity in their liver cytosol. Injecting pacus subjected to hypoxia during the cold season with a summer physiological dose of melatonin caused their liver cytosolic GST–HNE activity to increase up to the levels found in the warm season. From October to March (warm season in Pantanal), pacus are prone to oxidative stress particularly during potamodromous active oxygen-demanding swimming, when they migrate up rivers to spawn. Thus, our findings point out that the higher levels of melatonin in circulation during the summer are important to avoid the increase of 4-HNE inside liver cells of this fish species.     

Bending Characteristics of Polymethylmethacrylate Columns,Connecting Bars of Carbon Fiber,Titanium, and Stainless Steel Used in External Skeletal Fixation and an Acrylic Interface

Objective: To (1) mechanically evaluate polymethylmethacrylate (PMMA) columns of various sizes and compare them to connecting bar materials (carbon fiber composite, titanium, stainless steel) and (2) compare the properties of an intact PMMA column to those of an acrylic interface. Study Design: Experimental mechanical study. Sample Population: Experiment 1: 6 groups of 6 specimens each; experiment 2: 2 groups of 12 specimens each. Methods: All specimens were tested in 3‐point bending. Stiffness, yield strength, and ultimate strength values were calculated for each specimen. Results: PMMA 1 columns (23.25 mm) compared favorably to titanium or stainless‐steel bars. PMMA 3 columns (30.15 mm) and carbon fiber bars had similar yield strength but PMMA 3 columns were less stiff than carbon fiber bars. PMMA 3 columns had lower bending modulus and a higher variability in their mechanical properties than PMMA 1 or PMMA 2 (25.64 mm) columns. Acrylic interface specimens were less strong but as stiff as intact specimens. Conclusion: An acrylic interface was easily created and had acceptable biomechanical characteristics. Clinical Relevance: PMMA 2 and PMMA 3 columns are not recommended for clinical use at this time. Further tests are needed to analyze an acrylic patch in a more clinically relevant loading model.