Multiple blood meals in Anopheles darlingi (Diptera: Culicidae)

Anopheles darlingi is an important vector of human malaria in the Amazon. Adult females of this mosquito species require a blood meal to develop eggs, preferring humans to other blood sources. Although gonotrophic concordance has been described as the norm for An. darlingi, here we report An. darlingi female mosquitoes taking two or more blood meals within their first gonotrophic cycle. Only half of field‐captured adult females fed one blood meal developed follicles to Christophers' stage V. This outcome is dependent on larval nutrition, as 88% of laboratory‐raised well‐nourished females completed the first gonotrophic cycle with only one blood meal, while less nourished females needed additional blood meals. Half of the field‐captured blood‐seeking An. darlingi females had follicles in intermediate (IIIa and IIIb) and final (V) stages of the gonotrophic cycle, supporting the conclusion that An. darlingi blood feed more than once during a gonotrophic cycle. Additionally, we observed females attempting to blood feed a second time during the same day. Additional studies of An. darlingi biting behavior are necessary to accurately estimate Plasmodium sp. entomologic inoculation rates throughout the An. darlingi vast geographical distribution.     

A method of isolating the collagen (I) alpha2 chain carboxytelopeptide for species identification in bone fragments

We present a novel method for the isolation and analysis of the bone collagen (I) α2 chain carboxytelopeptide as a species biomarker. Conventional methods for the analysis and sequencing of mixtures of proteins and peptides commonly involve using the protease trypsin to cleave proteins present in the sample. However, in the study of collagen, these methods result in very complex mixtures of peptides that are difficult to analyze and the acquired results are not reproducible. Here we use bacterial collagenase (from Clostridium histolyticum) for its ability to cleave the highly unusual Gly-Xaa-Yaa repeating sequence of collagen, where Xaa usually is Pro and Yaa often is Hyp. Followed by a simple isolation step using a reverse phase solid phase extraction cartridge, the α2 (I) chain carboxytelopeptide can be readily analyzed by matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) and the results can be used to distinguish between different species of origin.     

酶水解制备花生粕多肽工艺的优化

目的：优化研究花生粕制备多肽的工艺。方法：采用酶法水解提取多肽、用总蛋白试剂盒（TP）双缩脲比色法在540nm处测定其含量。结果：用木瓜蛋白酶水解花生粕蛋白得到多肽的最佳工艺条件为：加酶量6 300u/g原料、温度45℃、底物浓度10%、酶水解时间5h。     

The effects of rapeseed meal and legume seeds as substitutes for soybean meal on productivity and gastrointestinal function in rabbits

The aim of this study was to determine the effects of soybean meal (SBM) substitution by a mixture of rapeseed meal (RSM), white lupine seeds (WLS) and pea seeds (PS) on productivity, nutrient digestibility, nitrogen retention and gastrointestinal function in Hyplus rabbits. The Control diet (SBM₁₅) contained 15% SBM, whereas Diet SBM_7.5 contained 7.5% SBM, 5% RSM, 4% WLS and 3% PS. In Diet SBM_0, SBM was completely replaced by RSM, WLS and PS (10%, 8% and 6%, respectively). A production trial was performed on 90 Hyplus rabbits aged from 35 to 84 d (45 each sex; 953 ± 4.6 g). A digestion and balance trial was conducted on 30 rabbits. Additionally, several parameters of the gastrointestinal tracts from eight animals from each group were analysed, where special attention was paid to the enzymatic activity of microbiota and the short-chain fatty acids concentration in caecum and colon. The experimental diets did not cause significant differences regarding performance parameters evaluated in vivo and post-mortem, and in the nutrient and energy digestibility or nitrogen retention. The observed changes in the enzymatic activity of large gut microbiota, including the selective increase in secretion of glycoside hydrolases by bacterial cells, seem to be responsible for the unchanged growth performance of rabbits fed diets where SBM was substituted by a mixture of RSM, WLS and PS. The obtained results indicate that in rabbit diets SBM may be, partially or completely, successfully replaced by a feed mixture of RSM, WLS and PS.     

Development of nitrogen and methane losses in the first eight weeks of lactation in Holstein cows subjected to deficiency of utilisable crude protein under restrictive feeding conditions

Low-protein diets are increasingly being used in dairy cow nutrition to minimise noxious nitrogen (N) emissions. However, at parturition, the lower milk yield at that time may mask deficiency in dietary utilisable crude protein (uCP; equivalent to metabolisable protein). Under restrictive feeding conditions, farmers would limit the feed allowance to match the lower measured milk yield, thereby exacerbating the deficiency. The consequences for N emission intensity per kg milk yield and methane emissions are unknown. In this study, two diets were fed to nine Holstein cows each from parturition onwards. One diet was complete and the other was calculated as 20% deficient in uCP. Feed allowance was always oriented towards the measured milk yield. In each of the first eight lactation weeks, intake and excretion were measured for 5 d. On the last 2 d of this period, methane emission was measured in respiration chambers. The statistical model included treatment, week and interaction as effects. The real levels of uCP and energy supply across the 8 weeks were 33% and 15% below requirements, respectively, in the Deficient cows. In addition, the Deficient cows consumed 18% less dry matter (caused by substantial refusals in week 1, where energy supply was according to requirements) and produced 25% less milk (26 vs. 34 kg/d). Cows in both groups used dietary N with similar efficiency for milk protein synthesis and excreted similar proportions of the N ingested via urine and faeces. This resulted in both treatments having similar N emission intensities per kg milk N and similar urinary N as a proportion of total excreta N, suggesting a similar potential for gaseous N emissions from the manure per kg of milk. The Deficient cows emitted 22% less methane overall but had similar methane yield and emission intensity to the Controls. In conclusion, a reduction in crude protein intake immediately after parturition does not reduce N emission per unit of milk when associated with uCP deficiency.     

玉米芯、竹炭及油枯吸附-海藻酸钠包埋施氏假单胞菌PFS-4对二氯喹啉酸的降解

采用玉米芯、竹炭及油枯吸附-海藻酸钠包埋对分离到的施氏假单胞菌PFS-4进行复合固定.采用正交试验对固定化条件进行优化,研究了固定化菌剂及游离菌体对二氯喹啉酸的降解效果.结果表明: 固定化菌剂制备的最佳条件为:海藻酸钠质量分数为4%、吸附载体比例(玉米芯∶竹炭∶油枯)为1∶2∶1、CaCl₂质量分数为3%、交联时间4 h.固定化菌剂在温度为30 ℃、初始pH=7的条件下,经6 d培养后,对浓度为800 mg·L^-1的二氯喹啉酸降解率为91.4%,而游离菌体的降解率为72.8%.将游离菌体和固定化菌剂用于实际污水及土壤处理时,固定化菌剂对水中及土壤中二氯喹啉酸去除率仍能分别达到84.2%和74.3%.研究结果表明,载体及其联结方式对土壤中二氯喹啉酸去除产生显著影响,翻动频率与土壤中二氯喹啉酸的去除率呈显著正相关.因此,玉米芯、竹炭及油枯吸附-海藻酸钠复合固定施氏假单胞菌PFS-4对不良环境具有较好的缓冲性能,对二氯喹啉酸污染水体及土壤原位生态修复具有潜力.     

Antioxidant content of oil and defatted meal obtained from borage seeds by an enzymatic-aided cold pressing process

Polyphenols content (as catechin equivalents) and tocopherol content were determined in borage defatted meal and borage oil, respectively. In addition, antioxidant activity of extracts obtained from borage defatted meal was evaluated. A cold pressing process was used for the extraction of Borago officinalis oil, resulting in a defatted meal (by-product). Polyphenols from this defatted borage meal were extracted using several solvents. An extract containing highly soluble solids and phenolic compounds with antioxidant activity (as free radical-scavenging, DPPH) was obtained when methanol was used. The tocopherol content was higher in oil extracted by cold pressing than in oil extracted with petroleum ether as organic solvent. An enzymatic treatment was applied (45 °C, 20% moisture, 0.25% E/S ratio, 1:1 Olivex:Celluclast enzymatic mixture) previously to borage oil extraction, which improved the antioxidant content in the borage defatted meal by three-folds, as compared to the values obtained by a nonenzyme-aided process.     

A PCR method for detection of plant meals from the guts of insects

The feeding behaviour of insects is a difficult ecological interaction to study. To date, entomologists have used biochemical and molecular techniques to identify the meals of predatory insects. We present here a molecular approach to identifying the DNA of plant species in the insect gut using the ribulose bisphosphate carboxylase gene large subunit (rbcL). A reference collection of 23 plant species from the southern Jordan Valley, Israel, was genetically characterized and employed. Insects belonging to eight different families were collected in the field along with the plants upon which they were found. After collection and prior to analysis, these insects were isolated on the plants they were found upon in the laboratory. This was to ensure that the insects had only one plant meal in their gut, as multiple plant meals would require additional techniques like cloning. A blind study was performed, genetically confirming plant DNA to species level from the processed gut contents of the insects. All reference plant species could be differentiated using a 157 bp long fragment of the rbcL gene. Plant DNA was identifiable, and the meal of the respective insect was accurately determined in each case. Analyses using experimentally fed crickets, Gryllodes hebraeus, determined that plant DNA was still detectable by PCR up to 12 h post-ingestion. This research proposes the application of molecular techniques for the identification of herbivorous insect feeding behaviour to increase understanding of plant–insect interactions.     

Effects of meal-time feeding and protein restriction on walking ability and some bone and carcass properties in broilers

This experiment was designed to investigate the effects of meal-time feeding and protein restriction on performance, gait score (GS) and carcass and bone traits in broilers. A total of 420 1-day-old chicks were wing banded and randomly distributed into 21 pens with 20 chicks each. At 7 days of age, chicks were weighed and randomly assigned to one of the three treatments: (1) control (C) feed (23.02% crude protein (CP)) was available ad libitum; (2) meal-time feeding (MF); control feed was available from 0100 to 0900 h and from 1500 to 2300 h. Food was withdrawn from 0900 to 1500 h and whole wheat (10 g/bird per day) was dispersed on the floor from 7 to 21 days; and (3) low-protein (LP) diet (19.71% CP) was fed to the chicks from 7 to 21 days. All of the groups were fed ad libitum from 1 to 7 days of age and from 21 to 45 days of age with a standard commercial diet. Individual body weight was measured on days 7, 21 and 45. Feed consumption was measured from 7 to 21 days and from 21 to 45 days. Forty-two chicks were humanly slaughtered and eviscerated for bone evaluation, on days 21 and 45. Also carcass characteristics were determined on day 45. Control group body weight was significantly higher (P < 0.05) at 21 and 45 days of age than the MF and LP groups, which did not differ. Feed intake was reduced by meal-time feeding and LP diet (P < 0.01). Feed efficiency was the best in the MF group during the period of 21 to 45 days of age (P < 0.01). In the control group, shank was significantly longer than that of the LP group and tibia breaking strength was higher than that of the MF group at 21 days (P < 0.05). However, shank width, tibia wet weight and tibia mid-diaphysis ash percentage of the MF group were significantly lower than those of the C and LP groups at 21 days of age (P < 0.05). GS, shank and carcass and tibia bone traits on day 45 were not significant among groups. No compensatory growth and walking ability improvement were observed at 45 days of age for broilers fed with MF and LP between 7 and 21 days of age.     

Detecting and quantifying meat meal or meat and bone meal contamination in fishmeal by visible and near infrared reflectance spectra

The use of animal protein feeds such as meat meal or meat and bone meal (MMBM) play an important role in the feed manufacturing industry, but their safe and healthy use in animal feeds is of public concern in order to prevent the spread of bovine spongiform encephalopathy (BSE). The objective of the present work was to develop a technique using near infrared reflectance spectroscopy (NIRS) that would be suitable for detecting and quantifying contaminating levels of MMBM in fishmeal. To this end, a partial least squares (PLS) discriminant analysis and a modified partial least squares (MPLS) quantitative analysis, using visible and NIRS, were developed using a calibration set of 186 samples including 90 samples of pure fishmeal and 96 samples adulterated with MMBM at levels ranging from 10 to 320 g/kg. An external validation set, comprised of 39 pure samples and 54 adulterated samples, was used to validate the calibration model. A PLS discriminant analysis model developed with mathematic pretreatment 1,4,4,1, successfully detected fishmeal adulterated with MMBM. External validation indicated that all samples were discriminated correctly. A MPLS quantitative model, developed with mathematic pretreatment 1,4,4,1, also successfully predicted the MMBM in fishmeal with standard error of cross-validation (SECV) of 27.89 g/kg and ratio of the standard deviation of the validation set to the standard error of prediction (RPD) of 3.37. The calibration and validation results confirm that NIRS could provide the feed industry and inspection bodies with a rapid, non-destructive and non-invasive technique for the detection and quantification of MMBM in fishmeal.