The conserved surface M-protein SiMA of Streptococcus iniae is not effective as a cross-protective vaccine against differing capsular serotypes in farmed fish

Streptococcus iniae causes invasive infections in fresh and saltwater fish and occasional zoonoses. Vaccination against S. iniae is complicated by serotypic variation determined by capsular polysaccharide. A potential target for serologically cross-protective vaccines is the M-like protein SiMA, an essential virulence factor in S. iniae that is highly conserved amongst virulent strains. The present study determined how SiMA is regulated and investigated potential as a cross-protective vaccine for fish. Electrophoretic mobility shift suggested that SiMA is regulated by the multigene regulator Mgx via a binding site in the −35 region of the simA promoter. Moreover, expression of simA and mgx was highly correlated, with the highest level of simA and mgx expression during exponential growth under iron limitation (20-fold increase in relative expression compared to growth in Todd-Hewitt broth). Based on these results, a vaccination and challenge experiment was conducted in barramundi (Lates calcarifer) to determine whether SiMA is protective against S. iniae infection and cross-protective against a different capsular serotype. The challenge resulted in 60% mortality in control fish. Formalin-killed bacterins prepared from the challenge strain resulted in 100% protection, whereas bacterins prepared from a serotypically heterologous strain resulted in significantly reduced protection, even when culture conditions were manipulated to optimise SiMA expression. Moreover, recombinant SiMA protein was not protective against the challenge strain in spite of eliciting specific antibody response in vaccinated fish. Specific antibody did not increase oxidative activity or phagocytosis by barramundi macrophages. Indeed incubating S. iniae with antisera significantly reduced phagocytosis. Lack of specific-antibody mediated opsonisation in spite of 100% protection against challenge with the homologous vaccine suggests that other immune parameters result in protection of challenged fish.     

Fine-tuning banana Xanthomonas wilt control options over the past decade in East and Central Africa

Xanthomonas wilt, caused by Xanthomonas campestris pv. musacearum has, since 2001, become the most important and widespread disease of Musa in East and Central Africa. Over the past decade, new research findings and especially feedback from small-scale farmers have helped in fine-tuning Xanthomonas wilt control options. During the initial years of the Xanthomonas wilt epidemic in East Africa, the complete uprooting of diseased mats and the burning or burying of plant debris was advocated as part of a control package which included the use of clean garden tools and early removal of male buds to prevent insect vector transmission. Uprooting a complete mat (i.e. the mother plant and a varying number of lateral shoots) is understandably time-consuming and labour intensive and becomes very cumbersome when a large number of diseased mats have to be removed. Recent research findings suggest that Xcm bacteria do not colonize all lateral shoots (i.e. incomplete systemicity occurs) and even when present that this does not necessarily lead to symptom expression and disease. This led to a new control method whereby only the visibly diseased plants within a mat are cut at soil level. The underlying idea is that the continued removal of only the diseased plants in a field will reduce the inoculum level and will bring down disease incidence to an acceptable level. This method is less labour intensive and takes a short time compared to the removal of a complete mat. However, single diseased stem removal needs to go hand in hand with prevention of new infections that can occur through the use of contaminated garden tools or through insect vector transmission. Novel transgenic approaches are also discussed. This paper presents an overview of past and ongoing research towards the development of a more practical and less demanding control strategy for Xanthomonas wilt.     

A secreted serine-threonine kinase determines virulence in the eukaryotic pathogen Toxoplasma gondii

Toxoplasma gondii strains differ dramatically in virulence despite being genetically very similar. Genetic mapping revealed two closely adjacent quantitative trait loci on parasite chromosome VIIa that control the extreme virulence of the type I lineage. Positional cloning identified the candidate virulence gene ROP18, a highly polymorphic serine-threonine kinase that was secreted into the host cell during parasite invasion. Transfection of the virulent ROP18 allele into a nonpathogenic type III strain increased growth and enhanced mortality by 4 to 5 logs. These attributes of ROP18 required kinase activity, which revealed that secretion of effectors is a major component of parasite virulence.     

Ovsynch Plus protocol improves ovarian response in anovular Murrah buffaloes in low‐breeding season

The objective of the study was to evaluate the efficacy of ovarian response and pregnancy rate in anovular buffaloes following Ovsynch and Ovsynch Plus protocols. Buffaloes (n = 55) were divided into two groups: Ovsynch group (n = 26): GnRH (10 μg, GnRH1) on Day 0, PGF₂α (25 mg) on Day 7, GnRH (10 μg, GnRH2) on Day 9; Ovsynch Plus group (n = 29): 500 IU equine chorionic gonadotropin (eCG) 72 hr (day ?3) prior to Ovsynch protocol, followed by fixed timed artificial insemination (FTAI) 6 and 24 hr after GnRH2 injection in bot groups. Transrectal ultrasonography was performed daily, that is, from day 0 and ?3 in Ovsynch and Ovsynch Plus group, respectively for ovarian response and pregnancy diagnosis at day 30 post‐insemination. In Ovsynch Plus group, administration of eCG prior to GnRH1 increased (p < .001) the diameter (mm) of dominant follicle (DF) from 10.15 ± 0.26 to 12.23 ± 0.34 within 72 hr of treatment resulting higher ovulatory response to GnRH1. Ovulation after GnRH1 was higher (p < .01) in Ovsynch Plus group (96.6%) than Ovsynch group (61.5%). However, ovulation rate to GnRH2 was similar (p > .05) between groups (Ovsynch group: 76.9% vs. Ovsynch Plus group: 70.0%). Mean DF diameter (mm) that ovulated to both GnRHs was higher (p < .01) than non‐ovulated counterparts in both groups (Ovsynch group: 10.80 ± 0.27 vs. 8.47 ± 0.53; Ovsynch Plus group: 11.99 ± 0.24 vs. 9.5 ± 0.63). Pregnancy was established in buffaloes which responded to both GnRHs, irrespective of groups, being higher (p = .52) in Ovsynch Plus group (34.5%) than Ovsynch group (23.1%), though non‐significant. In summary, this study showed that eCG inclusion prior to Ovsynch regimen improves ovulatory response in anovular buffaloes during low‐breeding season.     

Design,Synthesis and Evaluation of New Marine Alkaloid-Derived Pentacyclic Structures with Anti-Tumoral Potency

This work describes the synthesis and biological evaluation of a new heterocyclic hybrid derived from the ellipticine and the marine alkaloid makaluvamine A. Pyridoquinoxalinedione 12 was obtained in seven steps with 6.5% overall yield. 12 and its intermediates 1–11 were evaluated for their in vitro cytotoxic activity against different cancer cell lines and tested for their inhibitory activity against the human DNA topoisomerase II. The analysis by electrophoresis shows that the pentacycle 12 inhibits the topoisomerase II like doxorubicine at 100 µM. Compound 9 was found to have an interesting profile, having a cytotoxicity of 15, 15, 15 and 10 μM against Caco-2, HCT-116, Pc-3 and NCI cell lines respectively, without any noticeable toxicity against human fibroblast.     

Changes in pectins and product consistency during the concentration of tomato juice to paste

Concentrating tomato juice to paste during the tomato season allows for preservation and long-term storage, but subsequent dilution for formulation of value-added products is known to result in a loss of consistency. To understand the reasons for this, samples of unconcentrated juice, processing intermediates, and concentrated paste were collected from an industrial processing plant during normal commercial production. All samples were diluted with water to 5 degrees Brix and then analyzed for consistency and pectin content. Whole juice consistency, measured with a Bostwick consistometer, decreased through the course of juice concentration, with the largest change occurring early in the process, as the juice was concentrated from 5 to 10 degrees Brix. This decrease in consistency occurred during the production of paste from both hot- and cold-break juices. The change in Bostwick value was correlated with a decrease in the precipitate weight ratio. The loss of consistency during commercial processing was not the direct result of water removal because a sample of this same 5 degrees Brix juice could be concentrated 2-fold in a vacuum oven and then diluted back to 5 degrees Brix with no change in consistency or precipitate ratio. Total pectin content did not change as the juice was concentrated to paste, but the proportion of the total pectin that was water soluble increased. The greatest increases in pectin solubility occurred during the hot break and late in the process where the evaporator temperature was the highest.     

Nutrient digestibility profile of premium (category III grade) animal protein by-products for temperate marine fish species (European sea bass, gilthead sea bream and turbot)

Three trials, with classical experimental designs for in vivo digestibility studies, were conducted to determine the apparent digestibility coefficient (ADC) of protein (ADCp), lipid (ADCl), energy (ADCe) and amino acids (AA) in selected animal by-products fed to European sea bass, Dicentrarchus labrax (Trial 1), gilthead sea bream, Sparus aurata (Trial 2), and turbot, Psetta maxima (Trial 3). In each trial, five experimental diets [including a reference diet (RD)] where fish meal (FM) was used as the sole protein source were fed ad libitum to the fish for a period of 4 weeks. Test diets were based on the FM RD and obtained by replacing 30% of the RD with a category III designated European animal by-products (fit for human consumption), namely: steam hydrolysed feather meal (HFM), enzyme-treated feather meal (EFM), poultry meat meal (PMM) and spray-dried haemoglobin meal (SDHM). Faecal material was collected using the 'Guelph system', and nutrient and energy digestibility coefficients were related to the measurement of chromic oxide (Cr₂O₃) incorporated into the diet at a rate of 0.5%. Without any exception, FM diets yielded the best digestibility values for all macro-nutrients and by all fish. Among the test ingredients, ADCp was consistently higher for PMM and SDHM in the three species (85.5%, 91.1% in sea bass; 79.2%, 82.8% in sea bream; and 78.4%, 74.8% in turbot). Conversely, ADCp of HFM and EFM were less efficiently digested (67.2%, 68.2% in sea bass; 21.5%, 21.7% in sea bream; and 46.6%, 36.0% in turbot). However, the novel processing method applied to feather meal did not considerably influence the digestibility of most of the nutrients in this feedstuff. The current investigation yielded valuable numerical ADC for EAA considered to be of prime importance in generating balanced diet formulations.     

Cerebrospinal Fluid Changes in Two Horses With Central Nervous System Nematodiasis (Micronema deletrix)

Two horses with cerebrospinal nematodiasis (Micronema deletrix) had signs similar to those of other neurologic diseases resulting from parasitic (fly larvae, protozoa, or other helminths) migration through the central nervous system (CNS). In one horse (horse 1), a 13-year-old Paso Fino stallion, the cerebrospinal fluid (CSF) was slightly xanthochromic (1+), with a pleocytosis (25 nucleated cells/microliter) and a normal protein level (69 mg/dl). A CSF differential cell count showed 15% neutrophils, 56% lymphocytes, 22% macrophages, 5% eosinophils, and 2% basophils. In the other horse (horse 2), a 19-year-old Tennessee Walking Horse stallion, the CSF was modestly xanthochromic (2+), with pleocytosis (81 nucleated cells/microliter) and a modestly elevated protein concentration (114 mg/dl). A CSF differential cell count showed 9% neutrophils, 41% lymphocytes, and 50% macrophages. The CSF changes were consistent with those described for equine protozoal myeloencephalitis and verminous encephalitis. The microscopic lesions in both brains contained multifocal areas of malacia and granulomatous inflammation. Meningeal vessels throughout the brain were greatly thickened and inflamed, and they contained parasites. The CSF changes were not specific and histopathologic examination was required for a definitive diagnosis.     

Dysautonomia in a family of German shorthaired pointers.

Four, 5-week-old German shorthaired pointers from a litter of five puppies were presented for evaluation of vomiting and diarrhea. The dam was euthanized for suspected dysautonomia, and the fifth puppy died 1 day prior to presentation. On presentation, two puppies had vomiting and diarrhea, and the other two appeared healthy. The results of testing supported the diagnosis of dysautonomia in the two affected puppies but not in the healthy puppies. On day 5 of hospitalization, one apparently healthy puppy developed vomiting and diarrhea and had supportive testing for dysautonomia. Dysautonomia was confirmed at necropsy in affected dogs.