The treatment of bovine dermatophilosis and its effect on some haematological and blood chemical parameters.

In this study, the authors evaluated parenteral treatment of zebu cattle, with naturally and experimentally induced bovine dermatophilosis, in western Sudan, using four different antibiotic treatments. In terms of recovery rate, weight gain, avoiding relapse and preventing death, gentamycin was found to be the most effective treatment, followed by a combination of penicillin and streptomycin and, finally, long-acting oxytetracycline. However, enrofloxacin was not successful. A significant improvement in the red blood cell count was noticed among cattle treated with penicillin-streptomycin (p = 0.021) and gentamycin (p = 0.029). All treated cattle, except those treated with enrofloxacin, showed a significant improvement in mean corpuscular haemoglobin concentration (p = 0.021); mean corpuscular volume (p = 0.021), and white blood cell count (p < 0.021). Significant improvements were observed among treated cattle in their total levels of protein, calcium (p = 0.021) and cholesterol (p < 0.05), when compared to untreated cattle infected with Dermatophilus congolensis. This study recommends gentamycin as a drug of choice for the parenteral treatment of dermatophilosis. Treatment was not only effective in early, mild cases but also useful among moderately and heavily affected cattle. According to the observations of the authors, when no intervention took place, the condition of moderately and heavily affected cattle deteriorated and/or resulted in death.

Comparative assays of the rpoB gene for identification of Mycobacterium tuberculosis isolated from patients in Sudan.

OBJECTIVES: To characterise mycobacterial clinical isolates based on amplification of the rpoB gene. SETTING: One hundred and thirty-five mycobacterial isolates cultured from suspected pulmonary tuberculosis (TB) patients were identified phenotypically. Molecular characterisation of the isolates was performed based on amplification of the rpoB gene, using duplex polymerase chain reaction (DPCR), PCR-restriction fragment length polymorphism (RFLP) and nested PCR-based sequence analysis techniques. RESULTS: The DPCR assay identified 129 of 135 (95.5%) clinical isolates as Mycobacterium tuberculosis complex species. Restriction enzyme analysis of the rpoB PCR product using Hind II identified 134 of the 135 (99.3%) isolates as M. tuberculosis complex, while nested PCR sequence analysis of the rpoB gene identified 133/133 examined isolates (100%) as M. tuberculosis species. No mycobacteria other than M. tuberculosis (MOTT) were detected among the studied isolates. CONCLUSION: DPCR, PCR/RFLP Hind II and nested PCR sequence analysis of the rpoB gene techniques showed comparable efficiency in the characterisation of Mycobacterium isolates. Nested PCR sequence analysis of the rpoB gene was superior to PCR/RFLP for characterisation of suspected M. tuberculosis isolates, while the DPCR technique showed less sensitivity. As PCR-RFLP requires less sophisticated laboratory facilities than nested PCR sequence analysis, it would be more appropriate to be adopted for accurate characterisation of mycobacteria in countries with a weak infrastructure.

Unusual manifestation of a concurrent demodectic and sarcoptic mange in a Zebu-Friesian cross-bred heifer.

We describe a rare case of a concurrent demodectic and sarcoptic mange in a 2-year-old heifer in Khartoum, Sudan. The lesions were massive lumps of granulomatous tumour-like dermatitis with thick, nodular folds mainly covering the head, neck and shoulders. Histopathological examination of the lesions revealed the presence of both Demodex bovis and Sarcoptes scabiei. The animal died regardless of the anti-parasitic treatment it received.

Pulmonary nocardiosis caused by Nocardia otitidiscaviarum in an adult asthmatic female patient: The presence of acid-fast branching filaments is always significant.

We report a case of pulmonary nocardiosis in a 41-year-old asthma patient. Chest radiographs showed bilateral air space and consolidations.Acid-fast branching filaments were demonstrated in sputum, and the grown organism was identified phenotypically and confirmed using16S rDNA sequencing (accession no. KX500116). The patient received a combination of medical treatments, but developed complications,which were managed over the next 3 months, after which she was discharged. Pulmonary nocardiosis should be considered in patientsundergoing steroid therapy or when a chronic infection does not respond to first-line treatment.

Chronic rhinofacial basidiobolomycosis caused by Basidiobolus ranarum: Report of a case from Aseer Region, Kingdom of Saudi Arabia.

We present a case of fungal sinusitis caused by Basidiobolus ranarum in a 22-year-old male patient with chronic rhinosinusitis in Aseer region, Kingdom of Saudi Arabia. The patient was admitted with nasal obstruction accompanied by itching, sneezing, rhinorrhea, epistaxis and recurrent headache. Axial computed tomography (CT) scan of the paranasal sinuses showed a clear left facial swelling chronic inflammation and granulomata. Basidiobolus ranarum fungus was isolated on Sabouraud dextrose agar from a biopsy specimen. The organism was characterized by flat, yellowish-grey, glabrous, becoming radially folded fungus that under the microscope showed broad vegetative hyaline hyphae that bear zygospores with protuberances. The patient made good recovery and was discharged home with no recurrences after receiving oral itraconazole and removal of the polyps surgically.

In vitro anti-yeast activity of chloramphenicol: A preliminary report.

Chloramphenicol is a bacteriostatic antimicrobial agent but its antifungal activity is not known. The present study aimed to investigate the activity of chloramphenicol against 30 representative yeasts. The antimicrobial assay of chloramphenicol (50mg/mL; 100mg/mL and 200mg/mL) was determined by the disc diffusion method using Mueller-Hinton agar against 30 representative yeast strains. Zone of inhibition was read after 48-72h incubation at 37°C and results were compared with some standard antifungal agents. Most of the tested yeasts (73.3%) showed inhibition zones (5 up to 35mm) to chloramphenicol impregnated discs (200mg/mL). Three out of the four tested Candida albicans as well as Candida famata, Candida glabrata, Candida haemolonei and Cryptococcus neoformans showed no inhibition zones to chloramphenicol (200mg/mL). Caspofungin acetate (50mg/mL) inhibited 83.3% of the strains; ketoconazole (200mg/mL) 70% and metronidazole 10%. Chloramphenicol discs: 50 and 100mg/mL showed less activity (6.7% and 36.7%, respectively) compared to the 200mg discs; whereas chloramphenicol (BBL; 3µg/mL) inhibited 13.3% of the strains. The anti-yeast activities of chloramphenicol were comparable to other known antifungal compounds. Moreover, it is cheap, has fewer side effects and its inclusions in selective fungal media such as Mycosel have to be questioned.

Rapid identification of streptomycetes by artificial neural network analysis of pyrolysis mass spectra.

An artificial neural network was trained to distinguish between three putatively novel species of Streptomyces using normalised, scaled prolysis mass spectra from three representative strains of each of the taxa, each sampled in triplicate. Once trained, the artificial neural network was challenged with spectral data from the original organisms, the 'training set', from additional members of the putative novel taxa and from over a hundred strains representing six other actinomycete genera. All of the streptomycetes were correctly identified but many of the other actinomycetes were mis-identified. A modified network topology was developed to recognise the mass spectral patterns of the non-streptomycete strains. The resultant neural network correctly identified the streptomycetes, whereas all of the remaining actinomycetes were recognised as unknown organisms. The improved artificial neural network provides a rapid, reliable and cost-effective method of identifying members of the three target streptomycete taxa.

Molecular analysis of clinical isolates of Mycobacterium tuberculosis collected from patients with persistent disease in the Khartoum region of Sudan.

SETTING: Patients with positive smears for acid-fast bacilli were enrolled at tuberculosis (TB) clinics in the Khartoum region of Sudan. OBJECTIVE: To identify the presence of drug resistant genotypes in M. tuberculosis isolates which are difficult to treat. METHODS: Genus specific PCR-SSCP was performed to confirm the presence of M. tuberculosis in clinical isolates. Genotypic drug resistance testing was performed by mutation analysis and spoligotyping was used to monitor transmission and to identify epidemic strains. RESULTS: Fifty (48%) of the original 105 samples were classified as M. tuberculosis. Four (4%) of the samples were typed as mycobacteria other than TB, while the remaining (n =50) samples were refractory to further molecular analysis. The fifty amplifiable M. tuberculosis samples were used for subsequent mutation analysis and typing. Mutations were identified in the genes conferring resistance to INH (kat G, 12%), RIF (rpoB, 8%), SM (r psL and rrs, 30%) and EMB (embB, 4%). Two of the samples (4%) had mutations in genes associated to both INH and RIF and can be classified as MDR-TB. Thirty-three percent (13/39) of the persistant tuberculosis cases (5/18 treatment failure; 5/14 relapse; 3/7 defaulter) had mutations accounting for drug resistance. A total of 27 different spoligotypes were identified from 49/50 samples. Twenty-nine (59%) of the isolates were grouped into one of seven clusters, while 20 (41%) showed unique patterns. One patient was infected with M. bovis. CONCLUSION: This is the first molecular approach to characterize clinical isolates of M. tuberculosis from Sudan. The results show that drug resistance is indeed a serious problem and it may compliment the efforts of the National Tuberculosis Programme to improve strategies to control this disease.

Utility of chloranil in assay of naphazoline, clemizole, penicillin G sodium, and piperazine.

A simple and sensitive spectrophotometric method is described for the assay of naphazoline, clemizole, penicillin G sodium, and piperazine. The method was based on the formation of a charge transfer complex between these drugs as n-donors and chloranil, the pi-acceptor. Conformity to Beer's law enabled the assay of dosage forms of these drugs. Compared with official methods, the results obtained were of equal accuracy. A more detailed investigation of th naphazoline-chloranil complex was made with respect to its composition, association constant, and free energy change.

Utility of iodine and 7,7,8,8-tetracyanoquinodimethane for determination of terfenadine.

Simple and sensitive spectrophotometric methods for the assay of terfenadine are described. The first is based on the reaction of terfenadine with iodine to give a molecular charge-transfer complex, the terfenadine acting as an n-electron donor and iodine as a sigma-electron acceptor. The second depends on the formation of a highly coloured stable radical anion between terfenadine and 7,7,8,8-tetracyanoquinodimethane (TCNQ) as a pi-electron acceptor. Beer's law is obeyed over the terfenadine concentration range 0.2-1.2 mg/100 ml. The proposed methods have been successfully applied to the analysis of commercial terfenadine tablets.

Colorimetric determination of two fenamates in capsule dosage form.

The colorimetric determination of mefenamic acid and flufenamic acid with potassium ferricyanide in sodium hydroxide medium is described. The orange product is measured at 464 nm. The molar absorptivities are 1.9 x 10(3) and 2.9 x 10(3) 1.mole(-1).cm(-1) for mefenamic acid and flufenamic acid, respectively. The method has been applied successfully to the determination of these drugs in capsules.

Utility of 2,3-dichloro-5,6-dicyano-p-benzoquinone in assay of codeine, emetine and pilocarpine.

A simple and sensitive spectrophotometric method for the assay of codeine, emetine and pilocarpine is described, based on the interaction of these drugs (as n-electron donors) with 2,3-dichloro-5,6-dicyano-p-benzoquinone (as pi -acceptor) to give a highly coloured radical anion which exhibits maximum absorption at 460 nm. Formation of the radical anion has been established by electron spin resonance measurements. Beer's law is obeyed for the alkaloids investigated. The assay results are in accord with pharmacopoeial assay results. The procedure is sufficiently sensitive to permit unit dose assay of the individual alkaloids in pharmaceutical formulations.

Determination of diclofenac sodium, flufenamic acid, indomethacin and ketoprofen by LC-APCI-MS.

A sensitive, selective and accurate high-performance liquid chromatography-mass spectrometry (LC-MS) assay for the determination of selected non-steroidal anti-inflammatory drugs (NSAIDs), namely diclofenac sodium (DIC), flufenamic acid (FLU), indomethacin (IND) and ketoprofen (KET), either individually or in mixtures, was developed. The examined drugs were injected onto Shim-pack GLC-CN column and were eluted with a mobile phase consisting of acetonitrile and 20 mM ammonium acetate solution (5:1 v/v)/pH 7.4 at a flow rate l ml min(-1). The mass spectrometer, operated in the single ion monitoring mode, was programmed to admit the negative ions [M-H] at m/z 295.9 (DIC), 280.1 (FLU), 355.8 (IND) and 252.9 (KET), respectively. The calibration curves were linear (r > or = 0.9993) over the concentration range 50-300 ng ml(-1) (FLU, DIC) and 100-500 ng ml(-1) (KET, IND) with detection limits of 0.5-4.0 ng. The mean predicted concentrations for the analytes were in the range -5.9 and 5.2% of the nominal concentrations. Within-day and between-day precision were in the range of 0.8-9.1% of the R.S.D. Mean recovery percentages of the individual compounds from laboratory-made mixtures and pharmaceutical formulations were (99.5-101.5%) and (100.6-102.2%), respectively.

Physico-chemical characterization of a new salt of ibuprofen.

A new salt of ibuprofen was prepared by reaction with t-butylamine; its formation was confirmed by IR and 1H-NMR spectroscopy. The salt was characterized by thermoanalytical, X-ray powder diffraction and solubility studies. The salt was found to be 1.5 times more soluble in water than was ibuprofen, with an enthalpy of solution of -8.84 kcal mol-1.

Bovine farcy: a clinico-pathological study of the disease and its aetiological agent.

An extensive survey for bovine farcy covering 14,192 animals was conducted in the Sudan. The incidence of the disease was 31.9 per cent in Western Sudan which is regarded as an endemic area. Animals were examined clinically and at post-mortem. The lesions involved the prescapular, femoral, parotid, retropharyngeal, submaxillary, inguinal and mediastinal lymph nodes. Some animals had farcy lesions in the udder and some developed the pulmonary form of the disease. The disease is slow, but progressive in nature and was more prevalent among adult cattle than calves. Mycobacterium farcinogenes was found to be the cause of bovine farcy in the Sudan. The morphological and biochemical characteristics, and the mycolic acid content of the isolated strains were found to be identical to the reference strains of M. farcinogenes and were quite different from reference strains of Nocardia farcinica from which they were easily distinguished by the presence of alpha-mycolate, alpha'-mycolate and epoxymycolate in M. farcinogenes. Hydrolysis of mycolic acids with tetrabutylammonium hydroxide and the precipitation with toluene and acetonitrile gave good results comparable with acid methonalysis and alkaline hydrolysis. Precipitin lines were detected in the sera of some farcy-infected cattle. Histopathological examination of farcy lesions revealed a severe granulomatous reaction characterized by lymphocyte, macrophage, epithelioid and giant cell infiltration as well as marked fibrous proliferation. These changes were indicative of a progressive disease of the delayed tuberculin hypersensitivity type.

High-performance liquid chromatography-mass spectrometric analysis of furosemide in plasma and its use in pharmacokinetic studies.

This study presents a rapid, specific and sensitive high-performance liquid chromatography-mass spectrometric (LC-MS) assay for the determination of furosemide in human plasma using diclofenac as an internal standard (IS). Both compounds were extracted from human plasma with ethyl acetate at pH 1 and were chromatographed using Shim-Pack GLC-CN column and a mobile phase consisting of acetonitrile and 20 mM ammonium acetate buffer solution pH 7, 4:1 (v/v) at a flow rate 1 ml min(-1). Furosemide and IS were detected by mass spectrometer operated in the negative single ion monitoring mode using APCI as an ionization process at m/z 329.2 and 294.1, respectively. The assay linearity of furosemide was confirmed over the range 50-2,000 ng ml(-1). Detection limit for furosemide in plasma was 10 ng ml(-1). The selected concentration range corresponds well with the plasma concentrations of furosemide for pharmacokinetic study. Intraday and interday relative standard deviations were 1.3-4.7 and 2.7-11.5%, respectively. The extraction recovery percentages of furosemide and IS from plasma were in the range 89.3-97.1%. The developed LC-MS procedure was applied for the determination of the pharmacokinetic parameters of furosemide after an oral administration of tablet formulation (40 mg) to two healthy male volunteers. The calculated parameters were in good agreement with the reported values.

Comparative LC-MS and HPLC analyses of selected antiepileptics and beta-blocking drugs.

A highly sensitive and specific assay procedure based on the combination of liquid chromatography and mass spectrometry (LC-MS) has been developed for the quantitative analysis of selected antiepileptics (carbamazepine and phenytoin) and beta-blocking drugs (acebutolol, atenolol, pindolol and propranolol) using APCI as an ionization process. The measured concentration range was 100-300 ng ml-1 for all drugs except phenytoin (0.5-1.5 micrograms ml-1). Analysis was based on direct injection of methanolic solutions of drugs into the mass spectrometer with the subsequent elution with a mobile phase consisting of methanol and 1% acetic acid solution (4:1) at a flow rate 1 ml min-1. The mass spectrometer was programmed to permit detection and determination of either fragment or molecular ions of carbamazepine, phenytoin, acebutolol, atenolol, pindolol and propranolol at m/e 194.3, 252.9, 337.2, 267.1, 249.1 and 260.1, respectively. The recorded chromatograms exhibited well-resolved peaks at retention times < 1 min. The peak area was correlated linearly to the drug concentration. Intraday precision gave relative standard deviations in the range 1.75-4.02%. Compared to HPLC, the described LC-MS was faster, more sensitive and specific. Unlike HPLC, LC-MS could be applied to analyze incompletely resolved mixtures. The absolute detection limits for LC-MS and HPLC were 0.2-0.5 and 10-25 ng, respectively. Recovery studies of the investigated compounds in pharmaceutical products using LC-MS and HPLC gave mean percentages of 97.5-102.0 and 98.4-103.3, respectively. Statistical analysis of the data using t- and F-tests showed insignificant differences between both methods for the analysis of carbamazepine, phenytoin, acebutolol and atenolol in pharmaceutical formulations. However, LC-MS gave more accurate results than HPLC for determination of pindolol in tablets. Propranolol could only be determined in tablets using LC-MS.

Synthesis, spectroscopic characterization, stability assessment and DNA-binding of new 2,6-piperidinedione derivatives.

This work reports on structural characterization of new antineoplaston (ANP) representatives, namely 3-(benzoylamino)-2,6-piperidinedione (BPD), 3-(4-methoxybenzoylamino)-2,6-piperidinedione (MPD) and 3-(p-nitrobenzoylamino)-2,6-piperidinedione (NPD). These compounds were prepared by reacting N-(4-substituted benzoyl)-glutamines with N-hydroxysuccinimide to afford the corresponding esters, which were heated to produce the corresponding 2,6-piperidinedione (PD) compounds. Non-destructive analytical procedures such as 1H NMR and NIR analyses confirmed the postulated chemical structures of these PD compounds. HPLC chromatograms at an ambient temperature or from solutions preheated at 30, 40 or 60 degrees C displayed only a single peak for each compound. Combination of heat with pH modification had virtually no effect on the obtained peaks, thus attesting to the stability and purity of these compounds. MS analysis displayed molecular mass ions indicative of BPD, MPD and NPD at m/z 233.4, 263.2 and 278.3, respectively. The fragmentation patterns using MS/MS analyses conformed to the structural and molecular formulae of the prepared compounds. Furthermore, preliminary biological assessments showed the capacity of these compounds to bind to the DNA. NPD, but not BMP or MPD, had a superior affinity to the DNA than the prototype ANP-A10.

Nocardia farcinica--a significant cause of mastitis in goats in Sudan.

Fifteen of 100 mastitic milk samples from goats suffering from mastitis were tentatively identified as members of the genus Nocardia on the basis of selected phenotypic and chemotaxonomic characteristics. Six of the 155 strains were confirmed as Nocardia farcinica by 16S rDNA gene sequencing and subsequent aligning with relevant actinomycetes found in electronic databases and 2 by other identification criteria. N. farcinica is a serious cause of mastitis with a significant prevalence (15%) among the examined goats. Efforts are needed to optimise and simplify isolation and identification methods.

Streptomyces sp., a cause of fistulous withers in donkeys.

Four of 10 donkeys, which showed lesions simulating fistulous withers, were examined clinically with the aim to cultivate and identify the causal agent. Aspiated purulent materials were subjected to bacteriological examination. The causal organisms were recovered in Tryptic Soya agar medium when incubated aerobically at 37 degrees C for up to 5 days. These organisms were found to be actinomycetes-like, Gam positive with stable branching filaments and to form heavy aerial hyphae on colony surface. The isolated organisms ere tentatively identified as Streptomyces sp. on the basis of morphological and cultural characteristics. The initial sequences analysis of the 16S rDNA gene conformed that one of the isolates (SD551) falls within the phylogenetic clade, which encompasses the genus Streptomyces. Studies are underway to further describe the disease and its causal agent. The report represents a good evidence to incriminate Streptomyces in the aetiology of the fistulous withers.