Diet of crab-eating fox (Cerdocyon thous) in two conservation units of the Amazon rainforest, Brazil / Dieta do cachorro-do-mato (Cerdocyon thous) em duas unidades de conservação da Floresta Amazônica, Brasil

Abstract The present study aimed to evaluate the diet of the free-living crab-eating fox by identifying the stomach contents of the 17 crab-eating foxes (Cerdocyon thous) roadkilled in two conservation units, both located in the Amazon rainforest. The food items were quantified by frequency of occurrence (FO) and percentage of occurrence (PO). The stomach contents were analysed for dry matter (DM), crude protein (CP), crude fibre (CF), ether extract (EE), and mineral matter (MM). Nitrogen-free extractives (NFE), metabolisable energy (ME) values, as well as the energy need for maintenance were estimated. The composition of the diet for the crab-eating fox presented 29 food items from the different taxonomic groups, with a greater diversity of items of animal origin (n=22), although the highest frequency of occurrence was gramineae (Poaceae) (41.18%). Among the items of animal origin, 21% were mammals, 18% reptiles, 10% amphibians, 9% invertebrates and 3% birds. A high content of CF (62.76%) were determined. Nitrogen-free extractive and dry matter averages were 5.91% and 141.82 kcal/100g, respectively. The average maintenance energy was 447.01 kcal/day. These findings suggesting that the crab-eating foxes have a generalist diet with an omnivorous diet in the Amazon basin, feeding on gramineae, fruits, insects, snakes, amphibians, birds and small mammals and have the same feeding habit that present in other Brazilian biomes.

Resumo Este estudo objetivou analisar a dieta do cachorro-do-mato, de vida livre, por meio da identificação do conteúdo estomacal de 17 Cerdocyon thous atropelados em duas unidades de conservação da Floresta Amazônica. O conteúdo estomacal foi analisado e os itens alimentares foram quantificados pela frequência de ocorrência (FO) e percentagem de ocorrência (PO). Também foram avaliados os teores de matéria seca (MS), proteína bruta (PB), fibra bruta (FB), extrato etéreo (EE) e, matéria mineral (MM). Foram estimados o extrativo não-nitrogenado (ENN), a energia metabolizável (EM) e a necessidade energética de manutenção. A composição da dieta do cachorro-do-mato apresentou 29 itens alimentares dos diferentes grupos taxonômicos, tendo uma maior diversidade de itens de origem animal (n=22), ainda que a maior FO tenha sido de gramíneas (Poaceae) (41.18%). Dentre os itens de origem animal, 21% eram mamíferos, 18% répteis, 10% anfíbios, 9% invertebrados e 3% aves. Determinou-se um alto teor de FB (62.76%). A média do ENN e da MS foi 5.91% e 141.82 kcal/100g, respectivamente. A média da energia de manutenção foi 447.01 kcal/dia. Estes achados sugerem que o cachorro-do-mato encontrado na Bacia Amazônica é um animal generalista com uma dieta onívora, se alimentando de gramíneas, frutos, insetos, serpentes, anfíbios, aves e pequenos mamíferos, portanto com o mesmo hábito alimentar relatado a esta espécie quando encontrada em outros biomas brasileiros.

Diet of crab-eating fox (Cerdocyon thous) in two conservation units of the Amazon rainforest, Brazil

Abstract The present study aimed to evaluate the diet of the free-living crab-eating fox by identifying the stomach contents of the 17 crab-eating foxes (Cerdocyon thous) roadkilled in two conservation units, both located in the Amazon rainforest. The food items were quantified by frequency of occurrence (FO) and percentage of occurrence (PO). The stomach contents were analysed for dry matter (DM), crude protein (CP), crude fibre (CF), ether extract (EE), and mineral matter (MM). Nitrogen-free extractives (NFE), metabolisable energy (ME) values, as well as the energy need for maintenance were estimated. The composition of the diet for the crab-eating fox presented 29 food items from the different taxonomic groups, with a greater diversity of items of animal origin (n=22), although the highest frequency of occurrence was gramineae (Poaceae) (41.18%). Among the items of animal origin, 21% were mammals, 18% reptiles, 10% amphibians, 9% invertebrates and 3% birds. A high content of CF (62.76%) were determined. Nitrogen-free extractive and dry matter averages were 5.91% and 141.82 kcal/100g, respectively. The average maintenance energy was 447.01 kcal/day. These findings suggesting that the crab-eating foxes have a generalist diet with an omnivorous diet in the Amazon basin, feeding on gramineae, fruits, insects, snakes, amphibians, birds and small mammals and have the same feeding habit that present in other Brazilian biomes.

Resumo Este estudo objetivou analisar a dieta do cachorro-do-mato, de vida livre, por meio da identificação do conteúdo estomacal de 17 Cerdocyon thous atropelados em duas unidades de conservação da Floresta Amazônica. O conteúdo estomacal foi analisado e os itens alimentares foram quantificados pela frequência de ocorrência (FO) e percentagem de ocorrência (PO). Também foram avaliados os teores de matéria seca (MS), proteína bruta (PB), fibra bruta (FB), extrato etéreo (EE) e, matéria mineral (MM). Foram estimados o extrativo não-nitrogenado (ENN), a energia metabolizável (EM) e a necessidade energética de manutenção. A composição da dieta do cachorro-do-mato apresentou 29 itens alimentares dos diferentes grupos taxonômicos, tendo uma maior diversidade de itens de origem animal (n=22), ainda que a maior FO tenha sido de gramíneas (Poaceae) (41.18%). Dentre os itens de origem animal, 21% eram mamíferos, 18% répteis, 10% anfíbios, 9% invertebrados e 3% aves. Determinou-se um alto teor de FB (62.76%). A média do ENN e da MS foi 5.91% e 141.82 kcal/100g, respectivamente. A média da energia de manutenção foi 447.01 kcal/dia. Estes achados sugerem que o cachorro-do-mato encontrado na Bacia Amazônica é um animal generalista com uma dieta onívora, se alimentando de gramíneas, frutos, insetos, serpentes, anfíbios, aves e pequenos mamíferos, portanto com o mesmo hábito alimentar relatado a esta espécie quando encontrada em outros biomas brasileiros.

Oswaldocruzia franciscoensis n. sp. (Nematoda: Molineidae) in Leptodactylus macrosternum Miranda-Ribeiro, 1926 (Anura: Leptodactylidae) from Caatinga morphoclimatic domain, Brazil: morphological and molecular characterisation.

A new species of Oswaldocruzia Travassos, 1917 (Nematoda, Molineidae), parasite of Leptodactylus macrosternum Miranda-Ribeiro, 1926 (Anura: Leptodactylidae), from Caatinga morphoclimatic domain, Brazil, is described based on morphological and molecular data. Oswaldocruzia franciscoensis n. sp. is characterised by an anterior extremity with a cephalic vesicle divided into two portions, a body covered by cuticular longitudinal ridges, and cervical alae. Males of the new species have caudal bursa of type I with a 2-1-2 pattern, spicules divided into a shoe, bifurcated fork, and blade with two unequal branches, in which the longer branch bifurcates at its distal portion end and the smaller branch with three distal processes, each with distal bifurcations. Females have didelphic and amphidelphic uteri, an ovijector divided into vestibule, anterior and posterior sphincters, and anterior and posterior infundibula. The new species differs from its Neotropical congeners that have caudal bursa of type I, based on the presence of cervical alae and by having a spicular blade distally divided into two unequal branches, with the longer branch bifurcating at its distal portion and smaller branch with three distal processes, each distally bifurcated. The partial 18S rDNA sequence generated for Oswaldocruzia franciscoensis n. sp. is the first of a representative belonging to this genus in the Neotropical region.

Identification of carboxymethyl (CM)-binding proteins derived from Lolium multiflorum pollen extract and antibody reactivity in Brazilian allergic patients.

Lolium multiflorum grass is the major pollen allergen source in the southern region of Brazil, but most of its allergens remain poorly characterized. The aim of this study was to investigate antibody reactivity to L. multiflorum crude and carboxymethyl-ligand extracts in allergic patients and healthy individuals. Ion exchange carboxymethyl (CM) chromatography (CM-Sepharose) was used to isolate proteins (S2) from L. multiflorum crude extract (S1), which were assessed by SDS-PAGE. S1- and S2-specific IgE and IgG4 levels were measured by ELISA using sera from 55 atopic and 16 non-atopic subjects. Reactive polypeptide bands in S1 and S2 were detected by immunoblotting, and the most prominent bands in S2 were analyzed by mass spectrometry (MS-MS). Similar IgE and IgG4 levels were observed to both S1 (IgE median absorbance: 1.22; IgG4 median absorbance: 0.68) and S2 (IgE median absorbance: 1.26; IgG4 median absorbance: 0.85) in atopic subjects. S1 and S2 had positive correlations for IgE and IgG4 (IgE: r=0.9567; IgG4: r=0.9229; P<0.0001) levels. Homology between S1 and S2 was confirmed by IgE (84%) and IgG4 (83%) inhibition. Immunoblotting revealed that the 29-32 kDa band was recognized by 100% of atopic subjects in both S1 and S2. MS-MS analysis identified similarity profile to groups 1 and 5 grass allergens. This study revealed that carboxymethyl-ligand fraction played an important role for pollen allergy diagnosis by containing clinically relevant allergens and constituted a promising candidate for allergen-specific immunotherapy.

Identification of carboxymethyl (CM)-binding proteins derived from Lolium multiflorum pollen extract and antibody reactivity in Brazilian allergic patients

Lolium multiflorum grass is the major pollen allergen source in the southern region of Brazil, but most of its allergens remain poorly characterized. The aim of this study was to investigate antibody reactivity to L. multiflorum crude and carboxymethyl-ligand extracts in allergic patients and healthy individuals. Ion exchange carboxymethyl (CM) chromatography (CM-Sepharose) was used to isolate proteins (S2) from L. multiflorum crude extract (S1), which were assessed by SDS-PAGE. S1- and S2-specific IgE and IgG4 levels were measured by ELISA using sera from 55 atopic and 16 non-atopic subjects. Reactive polypeptide bands in S1 and S2 were detected by immunoblotting, and the most prominent bands in S2 were analyzed by mass spectrometry (MS-MS). Similar IgE and IgG4 levels were observed to both S1 (IgE median absorbance: 1.22; IgG4 median absorbance: 0.68) and S2 (IgE median absorbance: 1.26; IgG4 median absorbance: 0.85) in atopic subjects. S1 and S2 had positive correlations for IgE and IgG4 (IgE: r=0.9567; IgG4: r=0.9229; P<0.0001) levels. Homology between S1 and S2 was confirmed by IgE (84%) and IgG4 (83%) inhibition. Immunoblotting revealed that the 29-32 kDa band was recognized by 100% of atopic subjects in both S1 and S2. MS-MS analysis identified similarity profile to groups 1 and 5 grass allergens. This study revealed that carboxymethyl-ligand fraction played an important role for pollen allergy diagnosis by containing clinically relevant allergens and constituted a promising candidate for allergen-specific immunotherapy.

Diet of crab-eating fox (Cerdocyon thous) in two conservation units of the Amazon rainforest, Brazil.

The present study aimed to evaluate the diet of the free-living crab-eating fox by identifying the stomach contents of the 17 crab-eating foxes (Cerdocyon thous) roadkilled in two conservation units, both located in the Amazon rainforest. The food items were quantified by frequency of occurrence (FO) and percentage of occurrence (PO). The stomach contents were analysed for dry matter (DM), crude protein (CP), crude fibre (CF), ether extract (EE), and mineral matter (MM). Nitrogen-free extractives (NFE), metabolisable energy (ME) values, as well as the energy need for maintenance were estimated. The composition of the diet for the crab-eating fox presented 29 food items from the different taxonomic groups, with a greater diversity of items of animal origin (n=22), although the highest frequency of occurrence was gramineae (Poaceae) (41.18%). Among the items of animal origin, 21% were mammals, 18% reptiles, 10% amphibians, 9% invertebrates and 3% birds. A high content of CF (62.76%) were determined. Nitrogen-free extractive and dry matter averages were 5.91% and 141.82 kcal/100g, respectively. The average maintenance energy was 447.01 kcal/day. These findings suggesting that the crab-eating foxes have a generalist diet with an omnivorous diet in the Amazon basin, feeding on gramineae, fruits, insects, snakes, amphibians, birds and small mammals and have the same feeding habit that present in other Brazilian biomes.

Hydrostatic pressure mimicking diurnal spinal movements maintains anabolic turnover in bovine nucleus pulposus cells in vitro.

Treatment strategies for progressive intervertebral-disc degeneration often alleviate pain and other symptoms. With the goal of developing strategies to promote the regeneration of the nucleus pulposus (NP), the present study tried to identify the biological effects of hydrostatic (HP) and osmotic pressures on NP cells. The study hypothesis was that a repetitive regimen of cyclic HP followed by constant HP in high-osmolality medium would increase anabolic molecules in NP cells. Bovine NP cells/clusters were enclosed within semi-permeable membrane pouches and incubated under a regimen of cyclic HP for 2 d followed by constant HP for 1 d, repeated 6 times over 18 d. NP cells showed a significantly increased expression of anabolic genes over time: aggrecan, chondroitin sulfate N-acetylgalactosaminyltransferase 1, hyaluronan synthase 2, collagen type 2 (p < 0.05). In addition, the expression of catabolic or degenerative genes (matrix metalloproteinase 13, collagen type 1) and cellular characteristic genes (proliferating cell nucleic antigen, E-cadherin) was suppressed. The amount of sulfated glycosaminoglycan increased significantly at day 18 compared to day 3 (p < 0.01). Immunostaining revealed deposition of extracellular-matrix molecules and localization of other specific molecules corresponding to their genetic expression. An improved understanding of how cells respond to physicochemical stresses will help to better treat the degenerating disc using either cell- or gene-based therapies as well as other potential matrix-enhancing therapies. Efforts to apply these tissue-engineering and regenerative-medicine strategies will need to consider these important physicochemical stresses that may have a major impact on the survivability of such treatments.

Added sugar and trans fatty acid intake and sedentary behavior were associated with excess total-body and central adiposity in children and adolescents with congenital heart disease.

BACKGROUND: Over the past three decades, the prevalence rate of overweight and obesity has increased in survivors with congenital heart disease, and little is known about the body composition and its association with clinical characteristics and lifestyle factors. OBJECTIVES: To evaluate excess total-body adiposity and central adiposity and, to describe associated factors. METHODS: Cross-sectional study with children and adolescents who underwent procedure to treat congenital heart disease, from January to July 2017. Sociodemographic and clinical characteristics, and lifestyle factors (dietary intake, physical activity, and sedentary behavior) were assessed. Adiposity was assessed using air-displacement plethysmography and waist circumference. Factors associated with excess total-body adiposity and central adiposity were analyzed using logistic regression models. RESULTS: Of 232 patients, 22.4% were identified with excess total-body adiposity and 24.6% with central adiposity. Significant factors positively associated with excess total-body adiposity were intake of added sugar and trans fatty acids, adjusted for confounding factors. Similarly, lifestyle factors were positively associated with central adiposity: intake of added sugar and trans fatty acids, sedentary behavior, and family history of obesity. CONCLUSIONS: Lifestyle factors were associated with excess total-body adiposity and central adiposity. Assessment of body composition and healthy-lifestyle counseling into outpatient care may be the key point to prevent obesity in children and adolescents with congenital heart disease.

The use of Platelet-rich Fibrin in the management of medication-related osteonecrosis of the jaw: A case series.

Medication related osteonecrosis of the jaw (MRONJ) is characterized by exposed necrotic bone in the maxillofacial region that persists for more than eight weeks in patients taking antiresorptive or antiangiogenic drugs for bone metastasis or osteoporosis. The management of such condition depends on several factors, among which the staging of MRONJ. Though, a specific gold standard treatment has not been established to date. The aim of this case series is to describe the outcome of surgical treatment of MRONJ with the adjunct of Platelet-rich Fibrin (PRF). Eleven patients under therapy with alendronate underwent surgical removal of necrotic bone and debridement, followed by placement of PRF membranes in the bone defect. The outcome of the surgical treatment was successful in all patients, in a follow-up range from 12 to 36 months. In the cases presented, the macroscopic evaluation showed excellent and fast soft tissue healing, with no recurrence of bone exposure and no signs of infections. PRF membranes were also effective for postsurgical pain control. The use of PRF may represent a valuable adjunct in the surgical management of MRONJ.

Small for gestational age very preterm infants present a higher risk of developing bronchopulmonary dysplasia.

INTRODUCTION: Several studies assessed the influence of a low birth weight on bronchopulmonary dysplasia (BPD), but not all could find a significant association. Our aim was to assess the association between low birth weight and BPD in preterm infants, prospectively recruited at 11 level III Portuguese neonatal centers. METHODS: Obstetrical and neonatal data on mothers and preterm infants with gestational ages between 24 and 30 weeks, born during 2015 and 2016 after a surveilled pregnancy, were analyzed. Neonates were considered small for gestational age (SGA) when their birthweight was below the 10th centile of Fenton's growth chats and BPD was defined as the dependency for oxygen therapy until 36 weeks of corrected age. Statistical analysis was performed using IBM SPSS® statistics 23 and a p-value <0.05 was considered statistically significant. RESULTS: Out of 614, a total of 494 preterm infants delivered from 410 women were enrolled in the study; 40 (8.0%) infants with SGA criteria. SGA were more often associated with a single pregnancy, had greater use of antenatal corticosteroids, increased prevalence of gestational hypertensive disorders, C-section, rupture of membranes below 18 hours, rate of intubation in the delivery room, use of surfactant treatment, oxygen therapy, mechanical ventilation need, BPD, cystic periventricular leukomalacia, nosocomial sepsis and pneumonia; had lower prevalence of chorioamnionitis, and lower Apgar scores. The multivariate analysis by logistic regression, adjusted for BPD risk factors revealed a significant association between SGA and BPD: OR = 5.2 [CI: 1.46-18.58]; p = 0.01. CONCLUSION: The results of this study increase the scientific evidence that SGA is an independent risk factor for BPD.

Diurnal behaviors and herd characteristics of dairy cows housed in a compost-bedded pack barn system under hot and humid conditions.

With international demand for production systems aimed at thermal comfort and animal welfare, milk producers have become increasingly interested in compost barns. However, doubts about the behavioral aspects of cows in tropical and subtropical climates remain, because the compost barn system offers a larger bed area per animal at a lower stocking rate. The objective of this study was to evaluate whether the diurnal behaviors, hygiene and lameness of crossbred dairy cows are influenced by different number of lactations when housed in a compost-bedded pack barn system under hot and humid conditions. Crossbred cows (Holstein and Jersey), which were divided into two treatments based on number of lactations (primiparous and multiparous cows), were evaluated during lactation (n=12). The study was conducted from September 2015 to February 2016 in a compost barn in the southwest region of the state of Paraná, Brazil. Daytime behavior was observed between milking schedules using focal observations with 0/1 sampling. Cow hygiene and lameness were evaluated using subjective scores ranging from 1 to 4 and 1 to 5, respectively. Animal behavior and welfare was analyzed using Bayesian inference with a mixed effects model. The probabilities of dyspnea and pushing behaviors were higher (P<0.05) in multiparous cows, and the probability increased during the hottest hours of the day. For both multiparous and primiparous cows, the agonistic behaviors of pushing, butting and chasing peaked during the afternoon. Eating behaviors had the highest likelihood values (0.8 at 0800 h). The cows preferred to remain lying down in the morning, while rest and standing rumination were preferred in the afternoon. Primiparous cows were cleaner than multiparous cows, and the hygiene score for this group was significantly lower (P<0.05). The hygiene and lameness scores for all animals were low, and the highest scores were 1 and 2. In conclusion, multiparous cows exhibited more frequent agonistic behaviors during the hottest hours of the day. Regarding hygiene and lameness scores, multiparous cows exhibited a higher degree of dirtiness compared with the primiparous cows.

Exploring oyster mushroom (Pleurotus ostreatus) substrate preparation by varying phase I composting time: changes in bacterial communities and physicochemical composition of biomass impacting mushroom yields.

AIMS: To investigate characterization of the bacterial community composition and functionality and their impact on substrate biodegradation as well as mushroom yield. METHODS AND RESULTS: Bacterial diversity, composition and functionality were accessed by DNA-derived analysis for a sugarcane straw-based substrate composted for either 5, 10 or 15 days. In addition, carbon and nitrogen losses, carbohydrate conversion and mushroom yields were measured for the different treatments. Changes were observed in the bacterial community diversity and composition after the process started, but not during the composting process itself. Following phase I, Acinetobacter sequences were recovered in high numbers, and selected genes associated with nitrogen metabolism and lignocellulose deconstruction were mapped. Substrate physicochemical composition showed elevated carbon and nitrogen losses after 10 and 15 days of phase I with reductions in mushroom yield. CONCLUSIONS: Acinetobacter species appear to play an important role in substrate degradation processes, and a 5-day phase I period showed a significant higher mushroom yield compared to composting for either 10 or 15 days. SIGNIFICANCE AND IMPACT OF THE STUDY: This study confers a better understanding of the bacterial community manipulation during the substrate preparation and their influence in substrate selectivity for the Pleurotus ostreatus cultivation.

Helminth Parasites of the Lizard Nothobachia ablephara (Gymnophthalmidae) in Caatinga Areas from the Sertão of Brazil.

The lizard Nothobachia ablephara is endemic to dune areas and sandy soils adjacent to the São Francisco River in semiarid northeastern Brazil. Forty-nine lizard specimens were collected in 2 Caatinga areas in the municipality of Petrolina in Pernambuco state. Three gastrointestinal helminth taxa were identified, the nematodes Parapharyngodon alvarengai and Physaloptera sp. and the cestode Oochoristica sp. Nothobachia ablephara showed low parasite richness, but high levels of infection by P. alvarengai. There were no significant differences between the parasitism rates of the 2 study areas or between male and female lizards. This is the first study on parasitism in N. ablephara, thereby increasing knowledge of parasite fauna that infect gymnophthalmid lizards in the Sertão of Brazil.

Tadpoles fed supplemented diet with probiotic bacterium isolated from the intestinal tract of bullfrog Lithobates catesbeianus: Haematology, cell activity and electron microscopy.

The aim of this study is to select and isolate autochthonous bacteria with probiotic potential for use in a supplemented diet for bullfrog tadpoles, Lithobates catesbeianus. A total of 20 strains of lactic acid bacteria were isolated. Nine out of these were used in the following in vitro assays: antagonism against pathogenic bacteria (ANT), antimicrobial activity from extracellular compounds (MIC), tolerance to bile salts (TBS), pH reduction, protease production, sensitivity to antimicrobial tetracycline, cell viability, growth rate and doubling time. Using these data was defined an ideotype (ideal strain) based on the best results. Distances were estimated with the Mahalanobis (D2) test, and the best candidates, presenting the shortest ideotype distances, were considered to be used. The best strain was found to be Lactobacillus plantarum because it presented 10.00 ± 0.50 mm of ANT against Aeromonas hydrophila, 3.99 ± 0.01 of MIC independent of pathogenic bacteria, 85.07 ± 0.01 of TBS, 4.20 ± 0.02 of final pH, 17.67 ± 1.15 of protease production, 13.50 ± 2.00 sensitivity to antimicrobial tetracycline, 9.36 ± 0.04 of cell viability, 0.20 ± 0.00 of growth rate and 3.46 ± 0.00 doubling time. Therefore this probiotic candidate was then supplemented (2.045 ± 1.07 × 107 colony forming unities. g-1) into the diets of bullfrog tadpoles for a period of 42 days. At the end of the trial, samples of blood and intestines were collected to verify the haematological alterations and the intestinal morphology using transmission and scanning electron microscopy. Tadpoles fed the supplemented diet showed successful lactic acid bacterium colonisation, an increased number of circulating thrombocytes, monocytes, eosinophil and LG-PAS+ and also an increase in the length and density of intestinal microvilli. This study shows the feasibility of using probiotics isolated from farmed bullfrogs as a supplement in the diets of tadpoles, providing a promising alternative for modulating the health of these animals.

Dairy production practices and associated risks for bovine vaccinia exposure in cattle, Brazil.

A cross-sectional serosurvey was performed to identify environmental features or practices of dairy farms associated with risk for exposure to vaccinia-like viruses in dairy cattle in Brazil. Sera from 103 cows from 18 farms in Minas Gerais state were examined for Orthopoxvirus-neutralizing antibodies. A database of 243 binary or multiple-selection categorical variables regarding the physical features and surrounding ecology of each property was obtained. Thirteen of 46 presumptive predictor variables were found to be significantly associated with Orthopoxvirus serostatus by univariate logistic regression methods. Use of teat sanitizer and having felids on the property were independently associated with virus exposure by multivariable analysis. Rodents have long been suspected of serving as maintenance reservoirs for vaccinia-like viruses in Brazil. Therefore, domestic felids are not only effective predators of small rodent pests, but also their urine can serve as a deterrent to rodent habitation in buildings such as stables and barns. These results corroborate previous evidence of the high significance of rodents in the Vaccinia virus transmission cycle, and they also raise questions regarding the common use of teat sanitizers in dairy production areas.

Mating system and intrapopulational genetic diversity of Copernicia prunifera (Arecaceae): a native palm from Brazilian semiarid.

The aim of this study was to investigate the mating system and genetic diversity of the palm Copernicia prunifera using inter-simple sequence repeat markers. We found that the C. prunifera has multiple inflorescences with hermaphroditic flowers and pollen viability of 62%. Outcrossing rates at the population level (N = 267) produced a multilocus outcrossing rate (tm) of 0.878 and a single-locus outcrossing rate (ts) of 0.738, indicating that C. prunifera has a mixed mating system that is preferentially allogamous. The rate of mating among relatives (tm - ts) was low, indicating limited outcrossing between closely related individuals. The fixation index between seed trees (F) was negative (0.200), suggesting an absence of inbreeding, while the correlation of selfing (rs) was high (0.914). The values of the diversity indices among adults and progenies did not differ statistically (HE = 0.319 and I = 0.470; HE = 0.337 and I = 0.505, respectively). In testing for the presence of genetic bottlenecks using the infinite allele model and the stepwise mutation model, we observed a reduction in the effective population, as well as a deficit in heterozygosity (P < 0.0001). The results of this study inform management strategies for the conservation and genetic improvement of the C. prunifera palm.

Anthropization as a determinant factor in the genetic structure of Copernicia prunifera (Arecaceae).

This study uses ISSR molecular markers to characterize the demographic pattern, and spatial genetic structure (SGS) at different life stages of development (cohorts) in a natural population of Copernicia prunifera in the Rio Grande do Norte State, Brazil. All individuals were sampled and georeferenced in a 0.55-ha plot. The demographic analyses showed a clustered pattern in the first-distance classes and a random or segregated pattern at higher distance classes for all cohorts. Among the three studied life stages, juveniles showed the greatest value for Nei's genetic diversity index (HE = 0.369), while the lowest was found among reproductive adults (HE = 0.341). Greater genetic differentiation was found within life stages (98.61%) than among stages (1.39%). The total population (N = 161) showed positive and significant kinship in the first-distance class (12.3 m). The juveniles showed significant kinship up to 10.5 m. Non-reproductive adults had a positive kinship in the first-distance class (11.0 m) and a random distribution of genotypes in the remaining classes, while reproductive adults showed a random spatial distribution of genotypes. Tests for genetic bottleneck showed that the number of loci with excess observed heterozygosity was greater than expected. The SGS results reflected the restricted seed dispersion of the species, and the genetic bottleneck reflected the reduction of genotypes as a result of the anthropization of C. prunifera natural environments.

A Xanthomonas citri subsp citri hypothetical protein related to virulence contains a non-functional HD domain and is implicated in flagellar motility.

Citrus canker, caused by the Gram-negative bacterium Xanthomonas citri subsp citri (Xac), severely affects most economically important citrus varieties worldwide. A previous study showed that disruption of the ORF XAC1201 from the Xac 306 strain by transposon Tn5 decreased bacterium virulence in the Rangpur lime host (Citrus limonia L. Osbeck). However, little is known regarding the possible function of the hypothetical protein XAC1201 and how it affects the virulence of Xac 306. Here, we confirmed that disruption of ORF XAC1201 reduces Xac 306 virulence in two different hosts, delaying the onset of typical symptoms. In silico analysis suggested that XAC1201 interacts with the flagellar proteins FliM and FliL, known to be an important factor for virulence. In fact, motility assays revealed that the XAC1201 mutant has a significant difference in motility compared to the wild-type Xac 306. Also, a 3-D structure model revealed modified cofactor binding sites and suggested that XAC1201 has a non-functional HD domain. This hypothesis was confirmed by enzymatic assays performed in purified, XAC1201 recombinant protein expressed in Escherichia coli, which revealed no significant activities previously associated with HD domains for the tested substrates. Thus, the role of the XAC1201 protein in Xac 306 virulence seems to be related to flagellar motility, although a non-classic role for the HD domain cannot be dismissed.