Molecular typing of Mycoplasma agalactiae: tracing European-wide genetic diversity and an endemic clonal population

Mycoplasma agalactiae causes chronic infections in small ruminants and remains endemic in many regions of the world, despite intensive and costly eradication programs. In this study, the innate genomic plasticity of M. agalactiae was exploited to design and assess a combination of molecular epidemiological tools to trace the pathogen in different geographic locations and to understand its emergence or re-emergence after eradication campaigns. For this purpose, two collections of M. agalactiae isolates, representing European outbreaks or localized endemic disease in a single region of France, were subjected to RFLP (Restriction Fragment Length Polymorphism) analyses using two sets of DNA probes (distributed across the genome and specific for the vpma gene locus), and a previously described VNTR (Variable Number Tandem Repeats) analysis. A combination of four genome-specific DNA probes and two VNTRs gave the highest discriminative power. Molecular typing revealed that, while isolates from diverse geographical origins fell into clearly different groups, the endemic disease repeatedly observed in the Western Pyrenees region over the past 30 years has been caused by a unique subtype of M. agalactiae. This indicates that the re-emergence of the pathogen after seemingly successful eradication programs is not due to the importation of exotic strains, but to the persistence of local reservoirs of infection.     

Fatty acid,amino acid,and trace mineral analyses of five weaning foods from Jos,Nigeria

Five plant-based weaning foods (WF) (Dietrend, Jot-M, Soy, Ang and Vic-T) locallyprepared in Jos, Nigeria were analyzed by gas-liquid chromatography,reverse-phase high performance liquid chromatography, and atomicemission spectrometry with inductively coupled plasma to determine theirfatty acid (FA), amino acid, and trace mineral contents, respectively.Results of these direct analyses were compared to expected values derivedfrom food composition tables prepared by the United States Department of Agriculture (USDA). Additionally, results were compared against recommendednutrient values, using breast milk as the standard for FA content andrecommended dietary allowances (RDA) for amino acid and mineral contents.The overall nutritional value of the five WF varied considerably and thequantities of particular nutrients determined by direct analysis differedmarkedly from those estimated using USDA food tables. Comparison of WFfatty acid composition relative to the RDA recommendations and a humanmilk standard revealed a much higher proportion of both linoleic (35–55wt%) and -linolenic acids (1%–7 wt%) relative to human milklipids (11%–12% and 0.8%–0.9% wt, respectively); however, the WFwere devoid of arachidonic acid and docosahexaenoic acid. Soy containedthe highest amounts of linoleic acid (59.7 mg/g) and -linolenicacid (7.46 mg/g) compared to the other four WF (10.2–41.0 and 0.35–3.18 mg/g, respectively). The linoleic acid/-linolenic acid ratio was within the recommended range (5:1 to 10:1) in only Jot-M (10:1)and Soy (8:1). Dietrend, Vic-T and Ang, containedlinoleic/-linolenic ratios of 12:1, 29:1, and 82:1, respectively.The Soy weaning food would provide the most protein (24.3 g/day), basedon an estimated daily intake of 65 g of weaning food by a normalsix-month-old infant, compared to Jot-M (11.9 g/day), Dietrend (11.7g/day), Ang (8.07 g/day) and Vic-T (7.26 g/day). The protein RDA forchildren up to 1 year of age is 13–14 g/day. Comparison of the mineralcontents of the WF to the RDAs for various minerals indicated that all fivewould provide suboptimal amounts of calcium (16 to 250 mg/day) andzinc (1.42 to 3.56 mg/day) compared to respective RDAs of400 mg/day and 5 mg/day.These data show that the Soy weaning food is an excellent source of linoleicacid and -linolenic acid, as well as being a good source of highquality protein. Jot-M and Dietrend provide useful amounts of the essentialFA; however, it is advisable to reevaluate the composition of Ang andVic-T to find ways to improve the linoleic/-linolenic ratio of eachand increase their total protein content. These results document theshortcomings of using published food composition tables based on foods inAmerica when devising weaning foods based on ingredients in another partof the world.     

Nutrient Content of Four Edible Wild Plants from West Africa

Non-cereal plant foods in the Western Sahel of Africa contribute significantly to the diets of local residents, especially during periods of grain shortages. In this paper, we analyze four such plant foods including diyan kwakwa (nut of coconut palm, Cocos nucifera L.), muricin giginya (young shoot of Borassus aethiopum), tsamiya biri (fruit of the tree, Tamarindus indica), and yari (a mixture of lichens, mainly Rimelia reticulate) that grows on ebony trees (Diospyros mespiliformis). They were analyzed for their content of amino acids, fatty acids, and minerals. Although diyan kwakwa contained the highest protein content (27.1%), its protein quality fell below the WHO standard in 3 of 8 essential amino acid categories. Yari and muricin giginya contained moderate levels of good quality protein. Only diyan kwakwa contained calorically significant amount of total fatty acid (24.7%); however, none of the plants contained useful amounts of the essential fatty acids, linoleic acid, or α-linolenic acid. All four plants contained useful amounts of zinc (> 12 μg/g dry weight), while yari contained the most calcium (14.7 mg/g dry weight) and iron (1.41 mg/g), and diyan kwakwa the most copper. All the four plant foods contained lesser amounts of magnesium, molybdenum, or selenium. These data indicate that the four plants contain useful amounts of various essential nutrients that could supplement the diets of populations inhabiting the Western Sahel.     

Nutrient content of the edible leaves of seven wild plants from Niger

Wild plants play an important role in the diet of the inhabitants of Niger. These plants tend to be drought-resistant and are gathered both in times of plenty as well as times of need. Used in everyday cooking, famine foods may be an important source of nutrients. The goal of this study was to investigate the nutritional role of wild plants in the nigérien diet. To this end, leaves of seven plants species were analyzed for their mineral, amino acid and fatty acid contents: Ximenia americana, Amaranthus viridus, Corchorus tridens, Hibiscus sabdarifa, Maerua crassifolia, Moringa oleifera, and Leptadenia hastata. Ximenia americana} contained large amounts of calcium. Large quantities of iron were present in Amaranthus viridus. All seven plants contained significant amounts of selenium and phosphorus. Corchorus tridens contained the most protein (19–25% dry weight), and its composition compared favorably to the World Health Organization's standard for essential amino acids. Moringa oleifera contained 17% protein and compared favorably with the WHO standard. Corchorus tridens contained the largest amounts of the two essential fatty acids linoleic and -linolenic acids. These results reinforce the growing awareness that wild edible plants of the Western Sahel can contribute useful amounts of essential nutrients, including amino acids, fatty acids and trace minerals, to human diets.     

Analysis of nutritional components of eight famine foods of the Republic of Niger

In the western Sahel, indigenous plants become important staples when cereal harvests are inadequate to support populations inhabiting that region of Africa. The purpose of this study was to assess the nutrient content of several of these edible wild plants. The leaves of the following seven plant foods were analyzed: Ziziphus mauritiana, Cerathotheca sesamoides, Moringa oleifera, Leptadenia hastata, Hibiscus sabdarifa, Amaranthus viridi, and Adansonia digitata. The fatty acid, vitamin E, carotenoid, selected mineral and amino acid contents of these plant foods were determined. These same analyses were performed on the fruit of the Adansonia digitata. In quantitative and qualitative terms, Amaranthus viridis was found to be an excellent source of protein. Its amino acid composition compared favorably to that of a World Health Organization (WHO) protein standard. It also contained considerable amounts of the two fatty acids that are essential in humans (linoleic and -linolenic) and a number of minerals including iron, magnesium, calcium and zinc. The leaves of Hibiscus sabdarifa contained an appreciable quantity of protein the composition of which was comparable to the WHO standard. The mineral content of the leaves of this plant was also exceptionally high; noteworthy was its high zinc content. H. sabdarifa also contained significant quantities of the two essential fatty acids. Ziziphus mauritiana was an excellent source of the essential fatty acid linoleic acid and several of the metals including iron, calcium, magnesium and zinc. Its content of other essential nutrients, however, was rather low. In general, Adansonia digitata leaves were nutritionally superior to the fruit of the tree; however, the fruit did contain useful quantities of potassium, phosphorus, zinc and -linolenic acid. The Leptadenia hastata leaves were an especially good source of lutein and -carotene. These data should be useful to the people who inhabit the western Sahel in helping them devise healthy diets during times when cereal staples are in short supply.     

Amino acid, mineral and fatty acid content of pumpkin seeds (Cucurbita spp) and Cyperus esculentus nuts in the Republic of Niger

Dried seeds and nuts are widely consumed by indigenous populations of the western Sahel, especially those who inhabit rural areas. In light of the need for quantitative information regarding the content of particular nutrients in these plant foods, we collected dried pumpkin (Cucurbita spp) seeds and nuts of Cyperus esculentus in the Republic of Niger and analyzed them for their content of essential amino acids, minerals and trace elements, and fatty acids.On a dry weight basis, pumpkin seed contained 58.8% protein and 29.8% fat. However, the lysine score of the protein was only 65% relative to the FAO/WHO protein standard. The pumpkin seed contained useful amounts of linoleic (92 microg/g dry weight) and the following elements (on a microg per g dry weight basis): potassium (5,790), magnesium (5,690), manganese (49.3), zinc (113), selenium (1.29), copper (15.4), chromium (2.84), and molybdenum (0.81), but low amounts of calcium and iron. Except for potassium (5,573 microg/g dry weight) and chromium (2.88 microg/g dry weight), the C. esculentis nuts contained much less of these same nutrients compared to pumpkin seeds. In conclusion, pumpkin seeds represent a useful source of many nutrients essential to humans. The data in this report should of practical value to public health officials in rural areas of sub-Saharan Africa.     

The trypsin inhibitor content of 61 wild edible plant foods of Niger

In the western Sahel and many other regions of sub-Saharan Africa,wild edible plants contribute significantly to human diets, notonly during periods when cereal staples are scarce, but also whenthey are readily available. Although there have been publishedreports regarding the nutrient contents of these plant foods,little attention has been devoted to their content of antinutrientssuch as calcium chelators and inhibitors of the pancreas-derivedproteases, trypsin and chymotrypsin, which are required for theefficient digestion and absorption of dietary proteins. In thisstudy, aqueous extracts of 61 different leaves, seeds, fruits andflowers of edible plants gathered in the Republic of Niger wereanalyzed for their content of trypsin inhibitory substances using-N-benzoyl-DL-arginine-p-nitroanilide as the substrate andbovine trypsin as the enzyme source. Twelve of these plant foodscontained more antitrypsin activity than soybeans (1.34–8.18 vs.1.32 g trypsin inhibited/mg dry weight). Boiling for 3 mindid not inactivate the antitrypsin activity in most of the plantextracts. These data confirm that more than half of the wildedible plant foods widely consumed by various populations whoinhabit the western Sahel contain significant quantities of heat-stable trypsin inhibitor that could possibly compromise thebioavailability of proteins present in the diets of these populations.     

Use of ketoprofen as an antipyretic in cats.

The antipyretic effectiveness of Ketoprofen, a non-steroidal anti-inflammatory drug, was investigated in pyretic cats with a variety of bacterial and viral infections. Cats were randomly assigned to receive either a broad-spectrum antibiotic or a broad-spectrum antibiotic plus ketoprofen. Body temperature was monitored 3 times daily, and attitude and appetite were evaluated once daily. The treatment groups were compared with respect to mean body temperatures, using a one-way analysis of variance. Mean temperatures were significantly different (P < 0.05) during the 4 and 8 h post-treatment observations, with a reduction to normal temperatures in the ketoprofen group compared with no change in the group treated with antibiotics alone. The antipyretic effect of ketoprofen was rapid and persisted for at least 8 h, but for less than 24 h. The overall recovery period from pyrexia, depression, and inappetance was also shorter in cats treated with antibiotics and ketoprofen (3 d) than in cats treated only with antibiotics (5 d). Ketoprofen was a useful adjunct in the treatment of pyretic cats.     

Molecular basis of Mycoplasma agalactiae pathogenicity

Compared to other bacterial pathogens, the current knowledge of the molecular basis of pathogenicity of mycoplasmas is limited, and their strategies of infection at the molecular and cellular level remain to be elucidated. Several studies in the past years have shown that pathogenic mycoplasmas are equipped with sophisticated genetic systems, which allow these agents to spontaneously change their surface antigenic make-up. It is implicated that these variable surface components provide the wall-less mycoplasmas with a means to avoid the host immune response and promote host colonization. In Mycoplasma (M.) agalactiae, the agent of "contagious agalactia" in sheep and goats, a pathogenicity island-like locus has recently been identified that contains six distinct but related genes which encode the major immunodominant membrane proteins, the so-called Vpmas. It was shown that these surface-associated proteins vary in expression at an unusual high frequency due to site-specific DNA rearrangements. The previous lack of tools to genetically manipulate M. agalactiae has hampered more refined studies to assess the exact function of Vpmas in M. agalactiae infection and disease. The recent successful introduction of foreign DNA into the M. agalactiae genome therefore represents an important breakthrough which sets up the basis for a variety of follow-up studies assessing the role of Vpmas in molecular pathogenesis.     

Nutrient Content of Carob Pod (<Emphasis Type="Italic">Ceratonia siliqua</Emphasis> L.) Flour Prepared Commercially and Domestically

Although the fruit of the carob tree (Ceratonia siliqua L. Fabaceae) is nutritious and widely available in Turkey, especially in West and South Anatolia, much remains to be learned about its nutrient composition. The main goal of our study was to determine if there are differences in the content of certain nutrients in commercially-prepared carob flour (CPCP) and domestic or home-prepared carob powder (HPCP). Sucrose was the main sugar in CPCP and HPCP. Total protein was 40% lower in CPCP than HPCP due mainly to decreases in the content of several essential amino acids. However, except for lysine in CPCP, HPCP and CPCP compared favourably to a WHO protein standard. There were large differences in terms of their content of the two essential fatty acids, linoleic and α-linolenic acid, and the linoleic acid/α-linolenic acid ratio was 3.6 for CPCP, and 6.1 for HPCP. Manganese and iron were 2.5-fold higher in HPCP than CPCP. This study demonstrates that carob flour prepared in either the household or industrially is a good source of many, but not all essential nutrients, and that commercial processing of carob fruit into flour seems to affect its content of several important nutrients.