Colletotrichum species causing anthracnose on avocado fruit in Mexico: Current status

Colletotrichum species cause anthracnose disease in tropical and subtropical fruit crops worldwide. Mexico is the main producer and exporter of avocado (Persea americana) globally and has yearly outbreaks of anthracnose on this crop. However, which specific Colletotrichum spp. cause these outbreaks in avocado-producing regions remain uncertain; thus, the objective of the present study was to identify the species responsible. A survey performed in six production regions of Mexico yielded 232 isolates, from which a subset of 104 strains was selected based on morphological characteristics and origin. This subset was sequenced and haplotypes were analysed in the gapdh partial gene. Finally, 31 strains were identified through multilocus phylogenetic analyses using the sequences of the internal transcribed spacer region and six loci. This study revealed the presence of two species previously reported in Mexico (C. karsti and C. godetiae), three novel records in Mexico (C. siamense, C. fioriniae, and C. cigarro), four novel records on avocado (C. chrysophilum, C. jiangxiense, C. tropicale, and C. nymphaeae), and two novel lineages (Colletotrichum sp. 1 and Colletotrichum sp. 2). C. siamense was the most prevalent, while the species reported for the first time on avocado, including the novel lineages, were the least prevalent. C. karsti was the most widespread (four regions), followed by C. siamense, C. jiangxiense, and C. chrysophilum (three regions). Pathogenicity tests showed that all species caused anthracnose on avocado fruit. These findings will be useful for improving the management of avocado anthracnose outbreaks in Mexico.     

Effect of olive-derived antioxidants (3,4-dihydroxyphenylethanol and 3,4 dihydroxyphenylglycol) on sperm motility and fertility in liquid ram sperm stored at 15°C or 5°C

The aim of the present study was to evaluate the effect of the addition of two olive oil-derived antioxidants, hydroxytyrosol (3,4-dihydroxyphenylethanol, HT) and 3,4-dihydroxyphenylglycol (DHPG), on ovine semen during liquid storage at 5°C and 15°C. Semen was collected, pooled, diluted and then divided into aliquots supplemented with different concentrations (5 μg/ml, 10 μg/ml, 50 μg/ml and 100 μg/ml) of HT, DHPG and a mixture (MIX) of both antioxidants. Sperm motility characteristics were assessed in the different samples at 0, 6, 24, 48, 72 and 96 hr after cooling, and a fertility trial was also conducted. The results showed that the antioxidant addition did not significantly improve total and progressive motility in ovine cooled sperm maintained at 15° or 5°C. However, in samples stored at 5°C, LIN (48, 72, 96 hr), STR (0 hr) and WOB (0, 48, 72, 96 hr) values significantly decreased in comparison with control treatment when high antioxidant concentrations were added (MIX100 or HT100). When samples were maintained at 15°C, MIX50 showed significantly higher VCL values than the control treatment after 6 hr cooling, and MIX100 showed significantly lower VCL values at 96 hr after cooling. According to the artificial insemination trial, no significant differences were observed when antioxidants were added. In conclusion, the use of HT and DHPG showed small impact in sperm motility and fertility was not affected (nor detrimentally nor positively) when insemination was carried out using antioxidant-supplemented liquid sperm.     

The effects of time and dose of pregnant mare serum gonadotropin (PMSG) on reproductive efficiency in hair sheep ewes

The aim of this study was to evaluate the effects of dose and application time of pregnant mare serum gonadotropin (PMSG) on reproductive performance of hair sheep ewes synchronized with fluorogesterone acetate (FGA) under tropical conditions of Northeastern Mexico. Ninety-nine hair ewes (63 Blackbelly and 36 Pelibuey) were treated with intravaginal sponges during 10 days. After insertion of FGA sponges, ewes were divided into four groups, and PMSG was injected intramuscularly at doses of 100, 200, and 400 IU. Relative to FGA sponge removal, PMSG was administrated at −48 h, −24 h, and at sponge removal. PMSG was not administered to the control group. Control ewes had similar (P > 0.05) lambing rate, fertility, and fecundity than those treated with 100 IU of PMSG, but lower (P < 0.05) percentages to these variables than those treated with 200 and 400 IU of PMSG. Time to estrus decreased linearly, and ovulation rate increased quadratically as PMSG dose increased (0 to 400 IU). Administration of PMSG before sponge removal increased (P < 0.01) response to estrus and decreased (P < 0.01) interval to estrus compared with control. Ovulation rate, lambing rate, fertility, and fecundity were not affected (P > 0.05) by administration time of PMSG. Both dose and time of PMSG application did not affect (P > 0.05) pregnancy rate, percentage of single and multiple lambing, and prolificacy. In conclusion, results show that the dose of 400 IU of PMSG administered before sponge withdrawal in an estrus synchronization protocol improved reproductive efficiency of hair sheep ewes.     

Prevalence of extended-spectrum beta-lactamase-producing Escherichia coli isolates in faecal samples of broilers

Seventy-six faecal samples were obtained from broilers at slaughterhouse level in Portugal. Samples were inoculated on cefotaxime-supplemented Levine agar plates. Cefotaxime-resistant Escherichia coli isolates were recovered from 32 samples (42.1%), obtaining a total of 34 E. coli isolates (one or two isolates per sample). Susceptibility to 16 antibiotics was studied by disk diffusion method, and 85% of the isolates presented a phenotype of multi-resistance that included antimicrobial agents of at least four different families. Extended-spectrum-beta-lactamases (ESBL) of the TEM and CTX-M groups were detected in 31 ESBL-positive E. coli isolates. Twenty-six isolates harboured the bla_TEM-52 gene and two of them also harboured bla_TEM-1b. The bla_CTX-M-14 gene was identified in three isolates (in association with bla_TEM-1b in one of them), and bla_CTX-M-32 was demonstrated in two additional isolates. Three of the 34 cefotaxime-resistant isolates (9%) did not produce ESBLs, and two of them presented mutations at positions −42 (C → T), −18 (G → A), −1 (C → T), and +58(C → T) of the promoter/attenuator region of ampC gene. tet(A) and/or tet(B) genes were detected in all 34 tetracycline-resistant isolates, aadA in all 26 streptomycin-resistant isolates; cmlA in 3 of 6 chloramphenicol-resistant isolates, and aac(3)-II or aac(3)-I + aac(3)-IV genes in all 4 gentamicin-resistant isolates. Different combinations of sul1, sul2 and sul3 genes were demonstrated among the 22 trimethoprim–sulfamethoxazole-resistant isolates. Amino acid changes in GyrA and ParC proteins were identified in all 18 ciprofloxacin-resistant isolates. The results of this study indicate that the intestinal tract of healthy poultry is a reservoir of ESBL-positive E. coli isolates.     

Isolation and identification of phase II enzyme inductors obtained from black Shawaya sorghum [Sorghum bicolor (L.) Moench] bran

The antioxidant capacity as oxygen radical absorbance capacity (ORAC) and in vitro chemopreventive effect of phytochemicals extracted, partitioned and isolated from the decorticated bran of black Shawaya sorghum (Sorghum bicolor L. Moench) was studied. The chemopreventive effect was evaluated as the induction effect of quinone reductase (QR) using murine hepatoma cells Hepa 1c1c7. The crude methanol extract with an antioxidant capacity of 3.7 μmol Trolox equivalent (TE)/mg was partitioned into 120 fractions with fast centrifugal partition chromatography (FCPC). Fractions with partition coefficients between 0.1 and 0.26 exerted more than 50% of the antioxidant activity of the original crude extract and contained the highest concentration of anthocyanins. No correlation was found for ferulic acid content and ORAC or chemoprevention. A derivative of 7-methoxyapigeninidin at a concentration of 1 μg/ml increased by 31% the QR activity while the original extract reached a maximum of 27% at 40 μg/ml.     

Immunization with Eimeria ninakohlyakimovae-live attenuated oocysts protect goat kids from clinical coccidiosis

Caprine coccidiosis, affecting mainly young goat kids around the weaning period, is worldwide the most important disease in the goat industry. Control of caprine coccidiosis is increasingly hampered by resistances developed against coccidiostatic drugs leading to an enhanced need for anticoccidial vaccines. In the current study we conducted an oral immunization trial with live attenuated sporulated Eimeria ninakohlyakimovae oocysts. Sporulated E. ninakohlyakimovae oocysts were attenuated by X-irradiation technique. The experimental design included a total of 18 goat kids divided into the following groups: (i) animals immunized with attenuated E. ninakohlyakimovae oocysts at 5 weeks of age and challenged 3 weeks later with non-irradiated homologous oocysts (group 1); (ii) animals infected with non-attenuated E. ninakohlyakimovae oocysts at 5 weeks of age and challenged 3 weeks later with non-attenuated homologous oocysts (group 2); (iii) animals primary-infected with untreated E. ninakohlyakimovae oocysts at 8 weeks of age (control of the challenge infection, group 3); (iv) non-infected control animals (group 4). Goat kids immunized with live attenuated E. ninakohlyakimovae oocysts (group 1) excreted significantly less oocysts in the faeces (95.3% reduction) than kids infected with non-attenuated ones (group 2). Furthermore, immunization with live but attenuated oocysts resulted in ameliorated clinical coccidiosis compared to goat kids infected with untreated oocysts (group 2) and resulted in equally reduced signs of coccidiosis after challenge infection compared to acquired immunity driven by non-attenuated oocysts. Overall, the present study demonstrates for the first time that live attenuated E. ninakohlyakimovae oocysts orally administered showed almost no pathogenicity but enough immunogenicity in terms of immunoprotection. Importantly, vaccinated animals still shed low amounts of oocysts, guaranteeing environmental contamination and consecutive booster infections to sustain ongoing immunity.     

Role of seed infection by the Ascochyta blight pathogen of dried pea (Mycosphaerella pinodes) in seedling emergence, early disease development and transmission of the disease to aerial plant parts

The role of infected seed in the epidemiology of Ascochyta blight of pea, caused by Mycosphaerella pinodes, was studied both under growth chamber and field conditions, using healthy seeds, naturally infected seeds and artificially infected seeds. Results suggest that infected seeds caused serious losses, as a result of poor germination and high transmission of the disease, to parts of the plants under soil level. Foot rot symptoms often caused the death of young seedlings. Losses were increased by low temperatures during the early stage of crop development. M. pinodes progressed from seeds to aerial parts of the plants, but no Ascochyta blight symptoms occurred, the disease remaining near to the basal parts of the plants as a foot rot symptom. This suggests that seeds cannot be regarded as a source of contamination in the epidemiology of the disease.     

Digestibility of Buffel grass (Cenchrus ciliaris)-based diets supplemented with four levels of Gliricidia sepium hay in hair sheep lambs

The objective of this experiment was to evaluate the effect of supplementing increasing levels of Gliricidia sepium hay (GS) with different levels of inclusion of Buffel grass (BG) hay on digestibility by hair sheep lambs (2.5 to 3.5 months of age). Eight male lambs were used in a replicated 4?×?4 Latin square design with 21-day experimental periods (n?=?4). Animals were fed with BG with different levels of GS: 100 % BG (T1, control), 90 % BG?+?10 % GS (T2), 80 % BG?+?20 % GS (T3), and 70 % BG?+?30 % GS (T4). Dry matter (DM), neutral detergent fiber (NDF), and acid detergent fiber (ADF) intakes were not affected by treatments. The intake of crude protein (CP), organic matter (OM), and gross energy was higher (P?<?0.05) in those lambs fed with T4 diet than control. NDF and ADF digestibilities were higher (P?<?0.05) in T1 than in the other treatments. CP digestibility was higher (P?<?0.05) in T4 compared to T1, T2, and T3. The study showed that inclusion of GS up to 30 % with BG in forage-based diets of sheep does not affect DM and fiber intake nor influence DM and OM digestibilities but it increases CP intake and reduces NDF and ADF digestibilities.     

Lipid droplet distribution of immature canine oocytes in relation to their size and the reproductive stage

This study investigated the distribution of lipid droplets (LD) in immature canine oocytes in relation to their size and the reproductive stage. Oocytes were collected from the ovaries of bitches at different estrous stages, divided according to their size (110‐120 µm; >120 µm), and stained with Nile Red to detect lipid droplet distribution. At the follicular phase most of the oocytes displayed a diffuse pattern of LD distribution, whereas at anestrus and luteal phase oocytes showed LD mainly in a peripheral/ perinuclear LD distribution. A significantly higher intensity of LD has been recorded in the oocytes > 120 µm compared to those of smaller size (110 ‐ 120 µm) at all stages of the estrous cycle. At follicular phase, oocytes > 120 µm displayed LD intensity similar to that of oocytes > 120 µm at luteal phase and higher compared to the oocytes of the other groups.     

Development of a Model to Predict Runoff Water Headloss Through Compost Filter Berms

As a part of a larger study designed to develop compost filter berms i.e., an unsupported mound of compost that intercepts surface runoff from a site, suitable for removing nutrients and sediment from agricultural operations, we evaluated the hydraulic behavior of compost buffers with different masses and particle size distributions subjected to varying water flow rates using flume experiments. For each berm combination, the maximum flow rate the compost berm could sustain before a structural failure occurs was determined for different compost particle sizes. These data were used to develop models to predict headloss through a compost berm as a function of the flow rate per unit width, the median particle size of the compost media, and the buffer dimensions. The developed models perform well with R² values ranging from 0.64 to 0.97 with the majority of values greater than 0.9. The effect of presence of sediments in the flow was also quantified. The results show that headloss increased by 1–2 cm as a result of sediment removal, ranging from 5 to 35%, taking up pore space in the compost berm.