Bovine tuberculosis in South Darfur State, Sudan: an abattoir study based on microscopy and molecular detection methods

Bovine tuberculosis (BTB) is a widespread zoonosis in developing countries but has received little attention in many sub-Saharan African countries including Sudan and particularly in some parts such as Darfur states. This study aimed to detect bovine tuberculosis among caseous materials of cattle slaughtered in abattoirs in South Darfur State, Sudan by using microscopic and PCR-based methods. The study was a cross-sectional abattoir-based study which examined a total of 6,680 bovine carcasses for caseous lesions in South Darfur State between 2007 and 2009. Collected specimens were examined for the presence of acid-fast bacilli (AFB) by using microscopic and culture techniques. Isolated mycobacteria were identified by selected conventional cultural and biochemical tests in comparison to a single tube multiplex PCR (m-PCR) assay which detect Mycobacterium bovis-specific 168-bp amplicons. Of the total 6,680 slaughtered cattle examined in South Darfur, 400 (6 %) showed caseations restricted to lymph nodes (86.8 %) or generalized (13.2 %). Bovine tuberculosis was diagnosed in 12 (0.18 %), bovine farcy in 59 (0.88 %), unidentified mycobacteria in 6 (0.09 %), and missed or contaminated cultures in 7 (0.1 %). Out of 18 cultures with nonbranching acid-fast rods, 12 amplified unique 168-bp sequence specific for M. bovis and subsequently confirmed as M. bovis. With the exception of the reference M. tuberculosis strains, none of the remaining AFB amplified the 337-bp amplicon specific for M. tuberculosis. It could be concluded that bovine tuberculosis is prevalent among cattle in South Darfur representing 4.5 % from all slaughtered cattle with caseous lesions. The study sustains microscopy as a useful and accessible technique for detecting AFB. m-PCR assay proved to be valuable for confirmation of BTB and its differentiation from other related mycobacteriosis, notably bovine farcy.     

Assessment of adrenocortical activity by non-invasive measurement of faecal cortisol metabolites in dromedary camels (Camelus dromedarius)

The aim of this study was to determine whether glucocorticoid production could be monitored non-invasively in dromedary camels by measuring faecal cortisol metabolites (FCMs). Five Sudanese dromedaries, two males and three females, were injected with a synthetic adrenocorticotropic hormone (ACTH) analogue. Blood samples were collected pre- and post-ACTH injection. Faeces were sampled after spontaneous defecation for five consecutive days (2 days before and 3 days after ACTH injection). Baseline plasma cortisol values ranged from 0.6 to 10.8 ng/ml in males and from 1.1 to 16.6 ng/ml in females, while peak values after ACTH injection were 10.9–41.9 in males and 10–42.2 ng/ml in females. Peak blood cortisol values were reached between 1.5 and 2.0 h after ACTH injection. The concentration of FCMs increased after ACTH injection in the faeces of both sexes, although steroid levels peaked earlier in males [24 h; (286.7–2,559.7 ng/g faeces)] than in females [36–48 h; (1,182.6–5,169.1 ng/g faeces)], reflecting increases of 3.1–8.3- and 4.3–8-fold above baseline levels. To detect chromatographic patterns of immunoreactive FCMs, faecal samples with high FCM concentrations from both sexes were pooled and subjected to reverse phase high performance liquid chromatography (RP-HPLC). RP-HPLC analysis revealed sex differences in the polarity of FCMs, with females showing more polar FCMs than males. We concluded that stimulation of adrenocortical activity by ACTH injection resulted in a measurable increase in blood cortisol that was reliably paralleled by increases in FCM levels. Thus, measurement of FCMs is a powerful tool for monitoring the adrenocortical responses of dromedaries to stressors in field conditions.     

Evaluation of in vitro and in vivo inhibitory effects of fusidic acid on Babesia and Theileria parasites

Fusidic acid known to has antibacterial, antifungal, and antimalarial activities. Fusidic acid blocks translation elongation factor G gene in Plasmodium falciparum. In the present study, the inhibitory effects of fusidic acid on the in vitro growth of bovine and equine Babesia parasites were evaluated. The inhibitory effect of fusidic acid on the in vivo growth of Babesia microti was also assessed. The in vitro growth of four Babesia species that were tested was significantly inhibited (P < 0.05) by micromolar concentrations of fusidic acid (IC₅₀ values = 144.8, 17.3, 33.3, and 56.25 μM for Babesia bovis, Babesia bigemina, Babesia caballi, and Theileria equi, respectively). Combinations of fusidic acid with diminazene aceturate synergistically potentiated its inhibitory effects in vitro on B. bovis and B. caballi. In B. microti-infected mice, fusidic acid caused significant (P < 0.05) inhibition of the growth of B. microti at the dose of 500 mg/kg BW relative to control group. These results indicate that fusidic acid might be incorporated in treatment of babesiosis.     

Cytotoxic,Cytostatic and HIV-1 PR Inhibitory Activities of the Soft Coral Litophyton arboreum

Bioassay-guided fractionation using different chromatographic and spectroscopic techniques in the analysis of the Red Sea soft coral Litophyton arboreum led to the isolation of nine compounds; sarcophytol M (1), alismol (2), 24-methylcholesta-5,24(28)-diene-3β-ol (3), 10-O-methyl alismoxide (4), alismoxide (5), (S)-chimyl alcohol (6), 7β-acetoxy-24-methylcholesta-5-24(28)-diene-3,19-diol (7), erythro-N-dodecanoyl-docosasphinga-(4E,8E)-dienine (8), and 24-methylcholesta-5,24(28)-diene-3β,7β,19-triol (9). Some of the isolated compounds demonstrated potent cytotoxic- and/or cytostatic activity against HeLa and U937 cancer cell lines and inhibitory activity against HIV-1 protease (PR). Compound 7 was strongly cytotoxic against HeLa cells (CC₅₀ 4.3 ± 0.75 µM), with selectivity index of SI 8.1, which was confirmed by real time cell electronic sensing (RT-CES). Compounds 2, 7, and 8 showed strong inhibitory activity against HIV-1 PR at IC₅₀s of 7.20 ± 0.7, 4.85 ± 0.18, and 4.80 ± 0.92 µM respectively. In silico docking of most compounds presented comparable scores to that of acetyl pepstatin, a known HIV-1 PR inhibitor. Interestingly, compound 8 showed potent HIV-1 PR inhibitory activity in the absence of cytotoxicity against the cell lines used. In addition, compounds 2 and 5 demonstrated cytostatic action in HeLa cells, revealing potential use in virostatic cocktails. Taken together, data presented here suggest Litophyton arboreum to contain promising compounds for further investigation against the diseases mentioned.     

Sonographic characteristics of goat testis on water bath based ultrasonography

Assessing the health of the testes in domestic animals is an important aspect of the breeding soundness examination and selection. The aim of the present study was to develop a simple method for scanning and to establish ultrasonographically the gross anatomic structures of the goat testes. Six adult male goats were examined to study the sonographic appearance of normal testes and epididymides using a water bath based ultrasound scanning technique. The ultrasonographic examinations were done using a 5–9 MHz/60 mm (7.5 MHz) linear-array transducer and a B-mode scanner. The ultrasonographic examination was performed in goats after standardizing the procedure on six testes collected from slaughter house. Results showed that in live goats when the probe was placed directly over the scrotum it gave distorted and unclear image. In water bath method the entire scrotum was dipped into a container filled with water and linear probe was used to observe the sonographic features of the testis. Each testis was viewed vertically, resulting in longitudinal image which was frozen, measured and printed through a thermal printer. The results of the ultrasonogram revealed that the testicular parenchyma was homogenous and moderately echogenic throughout. The diameter (mean±se) of the right and left testes was 4.47±0.14 and 4.42±0.07 cm respectively and no significant difference was observed between the testes. The mediastinum testis was a 1.50±0.22 cm wide linear structure of greater echogenicity than the testicular parenchyma when viewed in the transverse plane and nearly circular echogenic “spot” in the midline of the testis when viewed horizontally. The head and tail of the epididymides were easily identified on all the testes, but the epididymal body and ductus deferens were difficult to identify consistently. The tail of the epididymis was easily identified on the distal end of the testis with sonolucent tubules and appeared sonographically as a ‘peaked cap’ upon the testicular parenchyma. The diameter (mean±se) of the tail of right and left epididymis was 2.11±0.18 and 1.92±0.06 cm and no significant difference was observed between epididymides. The vascular pampiniform plexus (1.42±0.18 cm) was easily identified on the proximal end of the testes. The tunics of the testes appeared as a bright echogenic line. Inter-testicular septum appeared between testes as a hyperechoic line. It is concluded that ultrasonography permits a noninvasive evaluation of the internal structure of the scrotum and testes and water bath based sonographic examination may prove to be a valuable simple diagnostic methodology for evaluating physiopathologic conditions of goat testes and can be employed as a routine investigative method during breeding soundness and clinical examination.     

Development and Characterization of Calcium-Alginate Beads of Apigenin: In Vitro Antitumor,Antibacterial, and Antioxidant Activities

The objective of this work was to develop sustained-release Ca-alginate beads of apigenin using sodium alginate, a natural polysaccharide. Six batches were prepared by applying the ionotropic gelation technique, wherein calcium chloride was used as a crosslinking agent. The beads were evaluated for particle size, drug loading, percentage yield, and in vitro drug release. Particle size was found to decrease, and drug entrapment efficiency was enhanced with an increase in the polymer concentration. The dissolution study showed sustained drug release from the apigenin-loaded alginate beads with an increase in the polymer proportion. Based on the dissolution profiles, BD6 formulation was optimized and characterized for FTIR, DSC, XRD, and SEM, results of which indicated successful development of apigenin-loaded Ca alginate beads. MTT assay demonstrated a potential anticancer effect against the breast cancer MCF-7 cell lines. The antimicrobial activity exhibited effective inhibition in the bacterial and fungal growth rate. The DPPH measurement revealed that the formulation had substantial antioxidant activity, with EC50 value slightly lowered compared to pure apigenin. A stability study demonstrated that the BD6 was stable with similar (f2) drug release profiles in harsh condition. In conclusion, alginate-based beads could be used for sustaining the drug release of poorly water-soluble apigenin while also improving in vitro antitumor, antimicrobial, and antioxidant activity.     

Effects of defoliation on whole‐plant maize characteristics as forage and energy crop

Hail affects yield and quality of maize crops, and consequences also depend on the growth stage at which the injury occurred. Whole‐plant maize (WPM) silage is often used within the same farm for cattle feeding and biogas production. The present study aimed to verify the effects of hail damage, simulated by artificial defoliation, on yield and chemical and nutritional features, as well as on biochemical methane potential (BMP) of maize forage. In a randomized block design with three replicates, four defoliation levels (0%, 33%, 66% and 100% of leaf area removal respectively) have been applied at either the V12 (12th leaf), R1 (silking) or R3 (milk) stages for two consecutive years. WPM yield, chemical and nutritional features, and BMP were measured. Dry‐matter (DM) yield per hectare was progressively reduced (p < 0.001) with increasing levels of defoliation and with application at V12 in comparison with R1 or R3 (1.26 vs. 1.39 and 1.46 kg ha^?1 for V12 vs. R1 and R3; p < 0.003). Nutritive value and BMP per unit of product were less altered than dry‐matter yield per hectare by defoliation. Anticipating defoliation reduced net energy for lactation (5.26 vs. 5.46 MJ kg^?1 DM for V12 and R3 respectively; p = 0.02). Total defoliation resulted in an accumulation of nitrates (NO₃) compared to the other treatments (3.98 vs. 1.53 g NO₃ kg^?1 DM; p < 0.001). BMP was mainly reduced by early and complete defoliation. Equations were developed to estimate the effects of defoliation on yield, composition, and nutritive and energetic values of WPM.     

Linking monitoring and intervention for improved management of tigers in the Sundarbans of Bangladesh

Baseline data on distribution and abundance of tigers in the Sundarbans is required to identify problem areas and evaluate management strategies. This paper outlines a khal (creek) bank survey of track set frequency throughout the Bangladesh Sundarbans to aid formulation of a management-driven monitoring program. Three teams of two observers surveyed a total of 1 201 km of waterways throughout the Sundarbans, recording 1 338 tiger track sets. These sets became unrecognizable as tiger sign after a mean 10 days (range 6-14). Proportion of detectable sign recorded was 0.91. Mean (±standard error) sample unit track frequency was 1.12 ± 0.86 track sets/km of khal. The mean coefficient of variance in sample unit track rate, estimated by multiple counts of six sample units, was 0.21 (range 0.06-0.34). Track frequency generally increased from northeast to southwest. Four sample units (6%) had signs of reproduction, with a mean litter size of 1.75 ± 0.5. Monte Carlo simulation suggests a monitoring program of one complete survey every two years will have power of 0.8 (α = 0.2), to detect track frequency declines of ?19% and increases of ?17%. We recommend this monitoring scheme be implemented on the Indian side of the Sundarbans to provide a standard assessment of the tiger population and to form the basis for setting management objectives and evaluating transboundary conservation initiatives.     

Performance of pyrazosulfuron-ethyl in non-puddled transplanted rainy season rice and its residual effect on growth of the succeeding crop in rice-wheat cropping pattern

Abstract

A study was carried out in sandy clay loam textured soil of Mymensingh, Bangladesh to evaluate weed control efficiency of pre-emergence herbicide pyrazosulfuron-ethyl either alone or in sequential application with post-emergence herbicide in transplanted rainy season rice under non-puddled strip-tilled (NPST) field condition and also to examine the residual effect of those herbicides on germination and growth of the subsequently grown wheat crop. Five treatment combinations of pyrazosulfuron-ethyl were tested against one weedy check and one weed-free check. The study revealed that NPST rice field was mostly infested by grass and sedge weeds and herbicide treatments offered a wide range of control (above 50% to 95%) on all types of weeds. Application of pyrazosulfuron-ethyl followed by (fb) orthosulfamuron fb butachlor plus propanil provided the most effective and economic weed control over two years of the study. Moreover, micro-plot bioassay study claimed germination and growth of subsequently grown wheat were not adversely affected by herbicides that were applied in rice. Therefore, application of pyrazosulfuron-ethyl followed by post-emergence herbicide could be effective and economic to control weeds in NPST rice under rice-wheat system, but proper rate and time of application should strictly be followed.