Absorption-spectrophotometric determination of DNA (deoxyribonucleic acid) in milk ad modum Feulgen. Accuracy of analysis, specificity, correlation to content of cells

The Feulgen reaction is examined by absorption photometric measurements at 565 nm with correction for varying background absorption at 485 nm. This correction is done to improve the accuracy of measuring.Examinations of the accuracy of analysis for the Feulgen reaction and the direct microscopic cell counts show that the former is of the same order, when the cell count is made at a working factor of 17,000.The standard deviations expressed in percentage of the reaction value are greater the lower the reaction is, whereas the absolute standard deviation is reduced.It has been demonstrated that addition of a few (< 5) µg of DNA per ml milk can give Feulgen reaction. The coefficient of correlation between log added amount of DNA and log Feulgen reaction is 0.98.The coefficient of correlation between log Feulgen reaction and log cell content depends on the accuracy at which both determinations are made. In a determination of cell content at a working factor of 550, and several determinations of the Feulgen reaction the coefficient of correlation (r) is found to be 0.98. A cell content determination at a working factor of 20,000 and a single determination of Feulgen reaction on each milk sample yields r = 0.83.An examination of foremilk samples of the same cell content reveals on an average the highest reaction in mastitis-affected quarters which have been infected within the last 4 weeks. Quarters that have been infected for more than 4 weeks, on an average show the second highest reaction, whereas quarters of physiological cell number show the lowest reaction.The DNA-content in most cases is found to be too high compared with the number of cells counted microscopically. The DNA-content in centrifugated cells corresponds to the one calculated theoretically. The surplus DNA-content can be demonstrated in the cell-free skim milk fraction, probably originating from destroyed cells.The studies performed suggest that determination of the content of 2-deoxyribose in milk by means of the Feulgen reaction is a more correct measure of the cell content in the milk than is the microscopic cell count. Studies are being continued for illustration of these conditions.     

Analysis of the genotype-by-environment interaction of spring barley tested in the Nordic Region of Europe: Relationships among stability statistics for grain yield

Variation in agronomic and quality characteristics was investigated in 220 Nordic spring barley cultivars across distinct environments (6 locations during 3 years) in the Nordic Region of Europe. The objectives of this research were to determine the importance of the genotype by environment interaction in all characteristics evaluated and to establish the relationship among different stability statistics for grain yield. Combined analysis of variance across locations indicated that both environments and genotype by environment interactions influenced significantly the cultivar phenotypes for all characteristics, irrespective of their type (row number) or earliness. The first two interaction principal component axes of the additive-main-effects-and-multiplicative-interaction (AMMI) model accounted together between 35% and 75% of the total genotype-by-environment interaction for all characteristics. Grain yield was, on average, higher in 2-row than in 6-row cultivars, which were significantly earlier in heading and grain maturity than the former. However, in some of the most northern locations, 6-row barley cultivars significantly outyielded on average 2-row barley lines. The genotype by location interaction variance (σ² _GL) accounted by each genotype was significantly associated to the deviation from regression (Tai's λ) while the coefficient of regression (β) was significantly correlated to the IPCA1 and IPCA2 of the AMMI model in 2-row, 6-row and early barley cultivars. This revised version was published online in August 2006 with corrections to the Cover Date.     

Homing of Gyrodactylus salaris and G. derjavini (Monogenea) on different hosts and response post-attachment

In natural European waters, the congeneric monogeneans Gyrodactylus derjavini Mikailov, 1975 and G. salaris Malmberg, 1957 are primarily found on brown trout Salmo trutta L. and Atlantic salmon Salmo salar L., respectively. Interestingly, rainbow trout, Oncorhynchus mykiss (Walbaum), originating from North America, is as susceptible as brown trout to G. derjavini. However, the mechanisms involved in this host specificity are poorly understood but may include behavioural, mechanical and chemical factors affecting parasite attraction, attachment, feeding, reproduction and host responses. In the present laboratory work, this question has been studied. Detached parasites (either G. derjavini or G. salaris) were offered a choice in small aquaria between fry of rainbow trout, Atlantic salmon and carp Cyprinus carpio L. Within 48 hours more than 90% of G. derjavini colonised rainbow trout and left salmon almost uninfected. Some parasites were found on carp. During the same time span, more than 60% of G. salaris attached to salmon, the rest infected rainbow trout and none were found on carp. Following attachment, the parasites need appropriate stimuli to initiate feeding and reproduction but even such a successful specific colonisation can be followed by a host response. Both humoral and cellular elements have been suggested to participate in these reactions but in the present work it was demonstrated by immunoblotting and immunocytochemistry that no antibodies in host mucus and host plasma bound to any parasite structures or epitopes.     

The role of litter beetles as potential reservoir for Salmonella enterica and thermophilic Campylobacter spp. between broiler flocks

We evaluated the role of beetles infesting broiler chicken rearing facilities as potential reservoirs for Salmonella enterica infections between successive broiler flocks. In addition, their role as potential reservoirs for thermophilic Campylobacter spp. was also investigated. Fourteen broiler houses located at 11 different farms were included in the study. The houses were nonrandomly selected on the basis of their salmonella status; nine were persistently contaminated with salmonella whereas five were salmonella negative. For each broiler house, two consecutive broiler flocks (i.e., 28 broiler flocks in all) as well as beetles collected during both rotations of production and in the empty period (after cleaning and disinfection) between these flocks were monitored for the presence of salmonella. Examinations for the presence of campylobacter in the same sample materials were also performed. Beetles sampled during production were positive for salmonella or campylobacter or both. Furthermore, in one house, the occurrence of Salmonella indiana in two consecutive broiler flocks coincided with the presence of S. indiana-contaminated beetles in the empty period between the flocks. The genotype of the identified S. indiana was in all cases identical when analyzed by pulsed-field gel electrophoresis. However, our results also suggest that salmonella from beetles may not always be transmitted to the chickens and that beetles living in contaminated houses can remain free of infection. All cases of campylobacter-positive beetle samples were detected in connection with a positive chicken flock; in no case was campylobacter isolated from beetles taken from the empty period between rotations. Four beetle species were identified during this study. Alphitobius diaperinus was found in all houses and was relatively abundant in most. Typhaea stercorea and Ahasverus advena were found in eight and nine houses, respectively, and were abundant in most of these. Carcinops pumilio was found in small numbers in eight houses. No other insect species was identified. These investigations have shown that beetles in broiler houses infrequently are positive for salmonella. However, transmission of S. indiana between two consecutive broiler flocks can coincide with the presence of salmonella-contaminated beetles in the empty period, indicating that the beetles were the reservoir of S. indiana between the two flocks. Concerning campylobacter, the results suggest that beetles do not play a significant role as a reservoir of campylobacter from one rotation to the next.     

The serologic response to Salmonella enteritidis and Salmonella typhimurium in experimentally infected chickens,followed by an indirect lipopolysaccharide enzyme-linked immunosorbent assay and bacteriologic examinations through a one-year period

Three groups of 100 individually marked salmonella-free chickens were followed for a period of 53 wk. The chickens were infected as day olds by crop instillation of 10(8) colony-forming units: one group with Salmonella enteritidis and a second group with Salmonella typhimurium. A third group was kept uninfected as controls. The groups were monitored bacteriologically by examination of cloacal swabs and organs and serologically by examination of serum and egg yolk by a lipopolysaccharide enzyme-linked immunosorbent assay throughout the period. Within the first week, 100% of birds in both infected groups were excreting salmonella bacteria in the feces. However, the number of fecal excretors declined rapidly with time, down to 6% in 16 wk for S. typhimurium and down to a similar level within the first 8 wk for S. enteritidis. For the latter, relapses with up to 40% positive birds were observed at the onset of egg production. For both S. typhimurium and S. enteritidis, positive bacteriologic cultures were obtained by sampling from internal organs at the end of the experiment, more than 1 yr from the time of infection. At the age of 6-7 wk, 50% of the chickens in the two infected groups showed a measurable serologic response in serum samples. The response persisted throughout the study in both serum and egg yolk samples. The inclusion of serologic methods is a valuable additional tool in the detection of salmonella in poultry, but serology should be used in conjunction with bacteriologic methods in surveillance programs, in particular to detect flocks in early stages of infection before a measurable serologic response has been raised.     

Selective haddock (Melanogrammus aeglefinus) trawling: Avoiding cod (Gadus morhua) bycatch

The critical condition of the North Sea cod stocks has resulted in restrictions on not only cod, but also haddock and other species that are caught together with cod. Thus full exploitation of the haddock stock is unachievable unless cod can be excluded from the haddock catch. We designed a selective trawl based on the behavioral differences between haddock and cod as they enter a trawl, i.e., cod stay close to the seabed whereas haddock rise above it. The trawl's fishing line is raised ～60 cm above the seabed to allow cod to escape beneath the trawl while haddock are retained. To collect the escapees, three sampling bags were attached beneath the raised fishing line. The selective haddock trawl reduced the total catch of cod by 55% during the day and 82% at night, and 99% of the marketable haddock was caught during the day and 89% at night. Cod escape rates were highly length dependent: smaller cod escaped the trawl in greater numbers than did larger individuals. Whiting, saithe, lemon sole, and plaice were included in the analysis.     

Maple syrup urine disease in calves: a clinical, pathological and biochemical study

The clinical, pathological and biochemical findings of a study of 30 Poll Hereford, Hereford, Poll Hereford cross or Hereford cross calves affected with branched chain ketoacid dehydrogenase (BCKAD) complex deficiency or maple syrup urine disease (MSUD) are presented. In breeding studies, 6 of 21 calves from obligate heterozygote matings were affected with MSUD, suggesting the disease is inherited in an autosomal recessive manner. Calves were clinically affected from birth, but there were variations in the subsequent course of progressive deterioration of central nervous system function. Concentrations of the branched chain amino acids and keto acids were elevated in pre-suckle plasma and cerebellar water content was higher in affected calves. Activity of BCKAD complex was minimal in fibroblasts cultured from an affected calf. Spongiform encephalopathy and elevated ratios of the branched to straight chain amino acids in formalin fixed cerebral tissue were found in a stillborn foetus and a 3-month-old Hereford calf. These findings suggest the disease occurs prenatally and that a delayed form may exist.     

A phenetic analysis of morphological variation in Citrullus lanatus in Namibia

Morphological data recorded from field trials using Citrullus lanatus germplasm collected in Namibia were used to analyse and compare the various morphotypes of this species. The experiment comprised wild types and local landraces as well as commercial cultivars. Cluster analysis supported the indigenous classification system used in Namibia, in which Citrullus types are distinguished based on gross morphology, ecology and usage and grouped into seed, cooking and fresh-eating (watermelon) types. Commercial watermelon cultivars formed a distinct cluster. Wide variation was found within the local types whereas the genetic basis of the commercial type appears to be narrow. The commercial cultivars were most closely related to local watermelon types and more distantly related to the wild types, whereas the cooking melons form an intermediate group.     

Effects of management practices on yield and quality of milk from smallholder dairy units in urban and peri-urban Morogoro,Tanzania

A longitudinal study design was used to assess the management, chemical composition of cows’ milk and quantify the microbial load of raw milk produced at farm level. Data were collected between December 2010 and September 2011 in Morogoro municipality. Milk samples were collected once every month and analysed for butter fat (BF), crude protein (CP), total solids (TS) and solids non-fat (SNF). Total bacterial count (TBC) and coliform counts (CC) were normalized by log transformation. The average milk yield was 7.0 l/day and was not influenced by feeding systems and breeds. Dairy cows owned by people who had no regular income produced more milk than government employees and retired officers. Means of BF, TS, SNF and CP were similar in different feeding systems. Wet season had significantly higher TBC (5.9 log₁₀ cfu/ml) and CC (2.4 log₁₀ cfu/ml) but feeding systems had no effect. Stocking density influenced TBC but not CC. It can be concluded that dairy cows produced low milk yield and its quality was poor.