Histochemical alterations in the duodenum of goats experimentally infected with Paramphistomum cervi

Certain histochemical alterations in the different tunics of duodenum in kids were studied 20, 40, 60 and 80 days post-infection (DPI) with Paramphistomum cervi and the results compared with those of uninfected kids. There was a general reduction of polysaccharide complex substances and glycogen at 20 DPI. A marked decrease in polysaccharide complex substances and glycogen at 20 DPI. A marked decrease in polysaccharide complex substances and glycogen was especially discernible in the Brunner's gland and muscularis mucosa 20 DPI. Thereafter, these substances gradually increased and at 80 DPI this decrease was fully replenished. A slight reduction in mucin and protein content of the infected duodenal goblet cells was noticed at 20 DPI. It is suggested that juvenile P. cervi utilize host-tissue polysaccharide complex substances and glycogen for their growth and development during duodenal migration.     

A macro- and light microscopical study of the pathology of Calicophoron microbothrium infection in experimentally infected cattle

Twelve Tuli weaner steers aged 1 year were randomly subdivided into three groups of four animals and infected with different doses of Calicophoron microbothrium metacercariae. Each animal in Group I received a low dose (LD) of 5 000 metacercariae, Group II a medium dose (MD) of 15 000 metacercariae, Group III a high dose (HD) of 25 000 metacercariae and one additional animal was kept as an uninfected control (C). After infection, one animal from each group was slaughtered on Day 28, 42, 56 and 84 post infection (pi) and samples from the ileum, jejunum, duodenum, abomasum and the rumen were collected for histopathological and cytological examination. On Day 28 pi, the gross pathological lesions observed in the duodenum of the LD and the MD animals were similar and comprised duodenal thickening, corrugation, hyperaemia, petechiation and ulceration. In the HD animal the duodenal lesions were similar but more severe. The abomasal folds were severely oedematous in the MD group and nearly occluded the abomasal lumen. Moderate oedema of the abomasal folds was also present in the LD and HD animals. The gross pathological lesions regressed in all the infected groups with increasing age of infection and had disappeared completely by Day 56 pi. On Day 28 pi the histopathological lesions in the duodenum and jejunum of the LD and MD groups were similar, comprising subtotal villous atrophy, hyperplasia of Brunner's glands and Peyer's patches and moderate infiltration of eosinophils, mast cells and a few globule leukocytes, basophils and lymphocytes in the lamina propria. The HD group had total villous atrophy, severe hyperplasia and cystic dilatation of Brunner's glands, which had expanded to cover the entire submucosa. On Day 42 pi the histopathological lesions were still present in the MD and the HD groups comprising subtotal villous atrophy and hyperplasia of Brunner's glands. Heavy infiltrations of eosinophils, moderate amounts of mast cells and a few basophils, globule leukocytes and lymphocytes were still present in the lamina propria of all three groups. On Day 56 pi, a few glands were still cystic in the MD and the HD groups. Moderate cell infiltrations were still present in the lamina propria of all the three groups and by Day 84 pi complete regeneration had occurred in all animals.     

Maturation of Paramphistomum cervi in sheep in India

Histological studies of the trematode Paramphistomum cervi collected throughout the year from the rumen of sheep, revealed the occurrence of gravid worms from April to August and immature worms from September to March. The maturation of the gonads and the accessory sex glands i.e., the prostate-, Mehli's- and vitelline-gland began during the period March-April and showed functional peaks during the period July-August, which coincided with the availability of the intermediate host, Indoplanorbis exustus in nature. Maturation as well as the incidence of infection of P. cervi have also been discussed in relation to various ecological factors, i.e., rainfall and temperature.     

Use of monoclonal antibodies developed against chicken coccidia (Eimeria) to study invasion and development of Eimeria reichenowi in Florida sandhill cranes (Grus canadensis).

Eimeria gruis and Eimeria reichenowi are common coccidial parasites of a number of species of cranes. Until recently, little was known about either the site for invasion or the dynamics of early development of the crane coccidia because of the difficulty of identifying sporozoites and early developmental stages of these parasites by conventional staining methods. In the present study, monoclonal antibodies (MAbs) elicited against Eimeria spp. of chickens and turkeys were found to cross-react with sporozoites and developmental stages of E. reichenowi in the tissues of Florida sandhill cranes (Grus canadensis). With these Mabs, E. reichenowi sporozoites were found in specimens taken at 6 hr postinoculation (PI) from just proximal to Meckel's diverticulum in the jejunum to the ileocecal juncture. Fewer were found in the ceca and rectum and none in the duodenal loop. At 24 hr PI, there were markedly fewer sporozoites and their location had shifted to the duodenum. No stages were seen in intestinal cells at 5 days PI (DPI), but trophozoites had developed in the liver and spleen. At 10 DPI, sexual stages were detected in the intestine from the duodenal loop through Meckel's diverticulum but not in other organs. By 14 DPI, numerous developmental stages were detected in the intestine (ceca and jejunum), liver, and lungs but not in the heart, kidney, or brain. The number, location, and maturity of the stages in the ceca differed markedly from those in the jejunum.     

Seasonal trends of Paramphistomum cervi in Tabasco, Mexico

Thirty rumens were examined every month from May 1989 to April 1992 to establish seasonal trends of infection and their relationship to the climatic factors caused by Paramphistomum cervi in cattle. The cattle were chosen from three municipalities of the Sierra region in Tabasco: Jalapa, Tacotalpa and Teapa. P. cervi was found throughout the year with prevalence between 3.33 and 96.67%, and with a yearly average of 39.10%. The major period of infection was shown in Jalapa. However, the maximum mean intensity was found in Teapa. The prevalence of the infection in all three sites was similar, showing the regional trend of P. cervi infection. Cattle were infected mainly throughout the rainy and windy seasons, during summer, autumn and at the beginning of winter.     

The susceptibility of Lymnaeid snails to Paramphistomum cervi infections in Mexico

The occurrence of Paramphistomum cervi has been reported in Mexico, but its intermediate hosts have not been identified. Five species of snails of the genus Lymnaea, L. bulimoides, L. columella, L. cubensis, L. humilis and L. palustris, were collected in Mexico. Only three of the exposed species, L. palustris, L. cubensis and L. humilis, became infected with P. cervi. The first of these snail species was highly susceptible to infection and a higher number of the exposed snails shed cercariae. It is suggested that L, palustris may act as an important vector of P. cervi in Mexico.     

Epidemiological survey of Neospora caninum infection in dogs from Romania

Native cathepsin-L cysteine proteinase (28 kDa) was purified from the excretory secretory products of Fasciola gigantica and was used for sero-diagnosis of F. gigantica infection in buffaloes by Dot-enzyme-linked immunosorbent assay (Dot-ELISA). The test detected F. gigantica field infection in these animals with a sensitivity of ～ 90%. No specific IgG antibody binding was displayed by sera obtained from 76 buffaloes considered to be Fasciola and other parasite-free by microscopic examination of faeces and necropsy examination of liver, rumen and intestine. Additionally, sera from 156 Fasciola-free buffaloes, yet infected with Gigantocotyle explanatum, Paramphistomum epiclitum, Gastrothylax spp., Strongyloides papillosus and hydatid cyst were all negative, indicating that F. gigantica cathepsin-L cysteine proteinase does not cross-react with these helminth parasites in natural infection of the host. The data indicated that cathepsin-L cysteine proteinase based Dot-ELISA reached ～ 90% sensitivity and 100% specificity with relation to above parasites in the detection of bubaline fasciolosis. The present Dot-ELISA diagnostic assay is relevant to the field diagnosis of F. gigantica infection in buffaloes.     

Duodenal Brunner's gland adenoma causing chronic small intestinal obstruction in a dog

An eight-year-old, neutered, female English springer spaniel was presented with a 14-month history of vomiting, marked weight loss and lethargy, all of which were non-responsive to medical therapy. Abdominal radiographs and ultrasonography showed an intraluminal soft tissue mass extending from the pyloric antrum to the proximal duodenum. Two spherical masses both approximately 8 cm in diameter were removed via a duodenotomy. Histopathological examination showed the first mass to be a trichobezoar and the aboral mass to be a Brunner's gland adenoma. Surgery resulted in a complete resolution of the clinical signs.     

Effects of time of day and monensin on the size distribution of particles in digestive tract sites of heifers fed corn silage

Effects of time of day and dietary monensin in the distribution of size of digesta particles in different digestive tract sites and their intersite relationships were examined in six heifers (290 kg BW) with ruminal, duodenal and ileal cannulas given ad libitum access to corn silage, with or without 100 mg monensin.head-1.d-1, in a two-period crossover design. Ingestive masticate and digesta of corn silage were collected via esophageal, ruminal or intestinal cannulas. The distribution of particulate matter retained on sieves with apertures larger than 20 microm was determined by wet-sieving. The cumulative distribution of particulate matter on a series of sieves was regressed on retaining sieve aperture to estimate the sieve aperture that would retain 50% weight of the particulate matter (median retaining aperture, MRA). The MRA of masticate was 6,494 microm. The MRA of digesta particles decreased (P less than .05) from ventral rumen (1,847 microm) to dorsal rumen (1,797 microm) to duodenum (346 microm), but increased to the rectum (359 microm). The MRA was lower (P = .044) for the monensin treatment only in feces. The MRA of particulate matter in the dorsal and ventral rumen, duodenum and rectum all changed (P less than .05) over 24 h. An inverse pattern between the MRA of ruminal and duodenal digesta occurred, presumably the result of a nycterohemeral pattern of eating and ruminating activity. Across sampling times, an inverse relationship existed between MRA of ventral rumen and duodenal digesta. This relationship suggests that a ruminal digesta raft composed of larger particles (immediately following major meals) is more effective than a raft of smaller particles (prior to such meals) in preventing flux of large particles to the duodenum.     

青贮玉米和混合干草对山羊胃肠内环境参数的影响

以8只装有永久性瘤胃瘘管和十二指肠近端T型瘘管的徐淮白山羊为试验动物,分为2组,分别饲喂青贮玉米和混合干草,研究山羊瘤胃和十二指肠内环境的昼夜动态变化。结果表明:①混合干草组瘤胃和十二指肠的pH值均极显著(P<0.01)高于青贮玉米组,其日内动态变化都表现出采食后先下降后上升的趋势;②混合干草组瘤胃液氨态氮浓度极显著(P<0.01)高于青贮玉米组,而十二指肠食糜的氨态氮浓度极显著(P<0.01)低于青贮玉米组;③混合干草组瘤胃乙酸浓度显著(P<0.05)高于青贮玉米组,丙酸浓度差异不显著(P>0.05),青贮玉米组丁酸浓度极显著(P<0.01)高于混合干草组;十二指肠液乙酸、丙酸、丁酸浓度青贮玉米组均极显著(P<0.01)高于混合干草组。     

滩羊羔羊瘤胃和小肠菌群多样性的比较研究

目的 研究滩羊羔羊瘤胃和小肠菌群多样性的差异。方法 选取健康的断奶滩羊公羔,屠宰后采集瘤胃、十二指肠、空肠和回肠内容物,利用16S rDNA高通量测序技术分析瘤胃和小肠菌群结构及多样性。结果 十二指肠样品Chao1指数高于瘤胃、空肠和回肠,瘤胃样品Shannon指数和Simpson指数均高于其他部位,但差异不显著（P>0.05）。瘤胃液中拟杆菌门（Bacteroidetes）、螺旋菌门（Spirochaetae）、丝状杆菌门（Fibrobacteres）的相对丰度显著（P<0.05）高于十二指肠、空肠和回肠,十二指肠中广古菌门（Euryarchaeota）的相对丰度与瘤胃相比有升高的趋势（0.05<P<0.10）,空肠和回肠中厚壁菌门（Firmicutes）的相对丰度显著（P<0.05）高于十二指肠。瘤胃中的未鉴定菌属（Unidentified）、理研菌属_RC9（Rikenellaceae_RC9_gut_group）、密螺旋体属_2（Treponema_2）、瘤胃球菌属_NK4A214（Ruminococcaceae_NK4A214_group）、普雷沃菌属_UCG-001（Prevotellaceae_UCG-001）相对丰度显著（P<0.05）高于十二指肠、空肠和回肠,空肠中Family_ⅩⅢ_AD3011_group相对丰度显著（P<0.05）高于其他部位。结论 滩羊羔羊瘤胃中细菌的多样性更丰富,瘤胃微生物与十二指肠、空肠和回肠的细菌区系显著不同。     

Parasites of livestock in Asir Province, southwestern Saudi Arabia

Thirteen parasite species, three trematodes, five cestodes, one nematode, and four arthropods, have been reported in livestock in Asir, southwestern Saudi Arabia. The occurrence of Dicrocoelium dendriticum in imported sheep and Paramphistomum cervi in indigenous cattle is recorded for the first time in Saudi Arabia.     

Effects of Fusarium toxin-contaminated wheat grain on nutrient turnover, microbial protein synthesis and metabolism of deoxynivalenol and zearalenone in the rumen of dairy cows

The aim of the present study was to examine the effects of feeding Fusarium toxin-contaminated wheat to dairy cows on nutrient utilization in the rumen and on duodenal flow of deoxynivalenol (DON), zearalenone (ZON) and their metabolites. Six dairy cows fitted with a large rumen cannula and a simple T-shaped cannula at the proximal duodenum was used in two experiments. The experiments included a control period in which the uncontaminated control wheat was fed and a period in which the control wheat was replaced by the Fusarium toxin-contaminated wheat (8.05 and 7.15 mg DON/kg and 0.26 and 0.1 mg ZON/kg in Expts 1 and 2 respectively). The wheat portion of the daily ration amounted to 50% on a dry matter (DM) basis and rations were completed with hay or grass silage. Five of the six cows were non-lactating and the total daily DM-intake ranged between 4 and 12 kg. The pH-values and the concentration of volatile fatty acids in ruminal fluid were not significantly influenced by feeding the contaminated wheat. In contrast, the postprandial ammonia concentration was consistently higher when the mycotoxin-contaminated wheat was fed. Moreover, the flow of microbial protein and utilizable protein at the duodenum were reduced at the same time. The concentrations of DON and ZON and of their metabolites in freeze-dried duodenal digesta were either not detectable or negligible during the control periods whereas distinct concentrations were measured during the periods where the contaminated wheat was fed. DON was nearly completely metabolized to de-epoxy-DON and the flow at the duodenum ranged between 4% and 28% of DON-intake. The ZON metabolites alpha-zearalenol (ZOL) and beta-ZOL were recovered at the duodenum beside the parent toxin ZON. Their recovery as a percentage of ZON-intake ranged between 43% and 132%. In conclusion, feeding of Fusarium toxin-contaminated wheat altered the ruminal protein utilization. The question of whether this effect was a result of the mycotoxin being present in the rumen or of Fusarium growth-related structural (cell wall) changes of the wheat grain needs to be clarified. The low recovery of DON at the duodenum would indicate either a nearly complete degradation of the molecule in the rumen or an absorption by the mucosa of the rumen, whereas the higher ZON recovery would suggest a lower degradation of the parent toxin in the rumen and/or recovery of some bile-originating entero-hepatic cycling ZON/metabolites.     

Propionic acid disappearance from the foregut and small intestine of the beef steer

Studies were conducted to determine the disappearance of propionic acid from the rumen and across the small intestine. Four crossbred beef steers, fitted with ruminal, duodenal and ileal cannulas, were given pulsed-continuous infusions of a nonabsorbable liquid phase marker (CrEDTA) and buffered propionic acid to achieve ruminal propionate productions (RPP) of 142 (basal), 567 and 997 mmol/h. Liquid flows from the rumen, and at the duodenum and ileum (2.75, 3.57 and .92 liters/h, respectively) were not affected by RPP (P greater than .23), although significant differences existed between sites (P = .001), reflecting a net addition of liquid between the rumen and duodenum and a net removal of liquid within the small intestine. Propionate disappearance from the rumen was 40 to 57% irrespective of RPP, with the complement passing from the rumen. Of the propionate that exited the rumen, 93 to 97% disappeared prior to entering the duodenum. Overall removal of RPP prior to the duodenum was 97 to 99% irrespective of RPP. Passage of propionic acid at the duodenum and at the ileum did not differ (P greater than .76) across all RPP, and the overall mean passages did not differ from zero (P greater than .34); similar results were observed for other VFA. These observations indicate that essentially all RPP is removed prior to entering the small intestine. Additionally, net propionate disappearance across the small intestine was undetectable and likely biologically irrelevant. Therefore, VFA absorption does not appear to be a limiting biological process in the beef steer.     

Histochemistry of selected peptidases in small intestine mucosa in piglets experimentally infected with Isospora suis

In the small intestine mucosa of 24 gnotobiotical piglets experimentally infected the first day post partum with oocysts of the coccidium Isospora suis, the activities of dipeptidylpeptidase IV (EC.3.4.14.5.; DAP IV) and gamma-glutamyl transferase (EC.2.3.2.2.; GGT) in the microvillous zone of enterocytes were evaluated by scanning densitometry. The tissue of the small intestine in piglets infected with a dose of 100,000 oocysts of the coccidia of I. suis was examined in the period from the first till the eleventh day post infection (DPI). In the control piglets at the age of 2-5 days it was found that most of the DAP IV activity was located in the microvillous zone of the enterocytes of the middle jejunum, rear jejunum and ileum. The DAP IV activity of duodenum mucosa was lower; as compared with the activity in the mucosa of the jejunum and ileum it reached 53-57%. In the case of GGT activity, the highest density values of the reaction product were recorded in the microvillous zone of enterocytes of the duodenum and the whole jejunum, the lowest in the ileum mucosa (86-89%) of the activity found in the duodenum and jejunum). During the experimental infection the infected piglets had a significant deficit of both peptidases, especially DAP IV (the whole studied period). The development of GGT activity was slightly different with the onset of the marked decline of the enzyme activity only on the fifth DPI. The lower GGT activity persisted till the eighth DPI. The density of the GGT reaction product began to return to the normal on the ninth to eleventh DPI. No predisposition in the location of the deficit was observed in the peptidases studied during the infection. The decline of the activity of both enzymes influenced also the mucosa of all studied parts of the small intenstine. The difference lay in the relevance of lowering of the density of reaction product of DAP IV and GGT on other DPI and in the different intensities of the return of the activity to the physiological normal.     

The Use of a “Tt” Shaped Cannula for Collection of Duodenal Digesta in Sheep

Six sheep were equipped with both a rumen fistula and a duodenal cannula. The duodenal cannula was made of two ordinary T shape cannulas which were vulcanized into a double T cannula. Both the rumen and the duodenal cannulas were made of soft rubber.In two experiments six different diets were fed. In the first experiment the diets consisted mainly of ammonia-treated straw (NH3-straw) plus barley or sodium hydroxide-treated straw (NaOH-straw) plus concentrate containing different N-supplements. In the second experiment, the treated straw (either NH3-straw or NaOH-straw) was fed alone. The animals in the two experiments were fed at maintenance level.Duodenal digesta were collected for periods of 12 h by inflating a balloon in the distal flange of the cannula, using pieces of foam rubber. The volume of fluid leaving the rumen was measured using the marker Gr-EDTA. The effect of digesta removal on blood constituents was studied. Three to four observations on each ration were made.Digesta flow and dry matter entering the duodenum were higher with NH3-straw than NaOH-straw fed either alone or with concentrate. However, there was a considerable variation of which a large part was of individual nature.The volume of fluid reaching the duodenum was always lower than the volume leaving the rumen, indicating a net absorption of water in the omasum and abomasum. This was estimated to be from 10.1 to 11.6 1/24 h in Experiment 1 and from 0.3 to 1.4 1/24 h in Experiment 2 (51–59 and 5–31 % of the ruminai outflow, respectively).A significant increase in plasma K and Mg and decrease in P_i and a-amino N concentration were observed due to 12 h collection of duodenal digesta. However, plasma Na, Ca, glucose and urea nitrogen concentrations remained at their pre-collection levels.Post-mortem examination of duodenum in sheep slaughtered after six and 10 months revealed a normal gut with no sign of dilatation in the immediate vicinity of the cannula.     

Fragmentation and ruminal escape of particles as related to variations in voluntary intake, chewing behavior and extent of digestion of potentially digestible NDF in heifers

The distribution of particles of differing sizes in ruminal, duodenal and fecal samples, the efficiency of particle breakdown and ruminal escape and their relationships to voluntary intake, chewing behavior and extent of digestion of potentially digestible NDF were examined in six heifers (290 kg average BW) with ruminal, duodenal and ileal cannulas. Animals had ad libitum access to corn silage, with or without 100 mg monensin head-1.d-1, in a two-period crossover design. Variations in voluntary intake by individual animals were positively related to size of digesta particles (median retaining aperture, MRA) in the dorsal rumen (P = .07) and rectum (P = .08), but not to MRA of particulate matter from the ventral rumen and duodenum. No significant relationships existed among eating or ruminating activities and distribution of particles of differing sizes in digesta from any of the digestive sites. The MRA of particulate matter in duodenal and rectal digesta were negatively related (P = .08 and P = .10) to extent of digestion of potentially digestible NDF (PDNDF) at these sites. Voluntary intake was related positively to efficiency of ruminative degradation of digesta particles appearing at the duodenum (P = .09) and to duodenal DM digesta flow per opening of the reticulo-omasal orifice (ROO; P = .02), the latter being negatively related to extent of digestion of PDNDF in duodenal digesta (P = .09). These results suggest that animals with higher intake capability are more efficient ruminators and can partially override constraining factors of particle size and byoyancy and thereby achieve a larger amount of DM flowing per opening of the ROO.     

Reflux of bile acids from the duodenum into the rumen of cows with a reduced intestinal passage

The purpose of this study was to find out whether the concentration of bile acids in the rumen of healthy cows differs from that of cows with a reduced gastrointestinal passage. The investigations were performed on 277 heifers and cows of the Swiss Braunvieh, Simmental and Swiss Holstein breeds. Forty of these had traumatic reticuloperitonitis, 30 had right or left abomasal displacement, 14 had an ileus of the duodenum, 36 had an ileus of the jejunum and 54 had a caecal dilatation. The other 103 animals had disorders which did not affect the digestive system and were used as controls. The mean (+/- se) bile acid concentration in the rumen fluid of the control animals was 1.0 +/- 0.30 mumol/litre. The bile acid concentrations in the rumen fluid of the cows with a duodenal or jejunal ileus were significantly higher than those of the other groups, demonstrating that bile acids may reflux from the duodenum into the rumen of cows with an ileus in the small intestine.     

Mouse mastitis model of infection for antimicrobial compound efficacy studies against intracellular and extracellular forms of Staphylococcus aureus

Despite the general in vitro susceptibility of Staphylococcus aureus isolates that cause infectious bovine mastitis, the pathogen remains difficult to eradicate with the available antibiotics. The capacity to survive intracellularly has been proposed as a factor contributing to the persistence of S. aureus in the bovine mammary gland. The costs associated with the use of cows or goats to assess the in vivo efficacy of new antibacterial compounds constitute a major drawback. Therefore, in the present study, a mouse model of intramammary infection has been characterized for in vivo testing of new experimental drugs. An inoculum of 100 CFU of S. aureus per gland caused an important level of infection with minimal tissue damage as observed at 24 h post-inoculation. By microscopy, polymorphonuclear neutrophil cell infiltration of the infected mammary glands was observed to increase over time. At 12-24 h of infection, the pathogen was primarily found alive and dividing in neutrophils and occasionally within mammary epithelial cells. Intramuscular or intravenous injections of cephapirin at t = 0 and 10 h reduced the number of CFU/g of gland in a dose-dependent manner. In conclusion, the mouse model of infectious mastitis proposed here is suitable for primary evaluation of experimental drugs after parenteral treatment of intramammary infection with a pathogen such as S. aureus that presents both intracellular and extracellular phases of growth.     

Urinary purine derivates excretion as an indicator of in vivo microbial N flow in cattle: A review

Microbial protein flow to the duodenum may be regarded as the most important and sensitive indicator to optimise rumen metabolism in high-yielding dairy cows. In this review, the methodology and the sources of variation to estimate the duodenal microbial N flow with urinary excretion of purine derivatives (PD) as a non-invasive method is discussed. The urinary PD excretion was linearly related with the amount of purine bases (PB) infused in the abomasum or duodenum, but the recovery of PB in urine differed between experiments. The main sources of variation in the relationship between microbial N flow and urinary PD excretion are dietary contribution of nucleic acids to duodenal flow, varying N:purine ratio in duodenal digesta, differences in intestinal digestibility of nucleic acids and infused PB, and endogenous contribution of PD to urinary excretion. The recycling of PD to the rumen is negligible, and does not explain the incomplete urinary recovery of PD. A large proportion of the total PD is excreted as allantoin in urine. In some experiments this proportion was constant, whereas in others it varied with diet or physiological state of the animal. The excretion of PD in milk is not a suitable indicator of microbial N flow, due to mammary purine catabolism to uric acid and due to the strong positive correlation between milk allantoin excretion and milk yield. Instead of total urine collection, the molar ratio between urinary PD and creatinine can be used to estimate microbial N flow. However, a substantial between-animal variation in this ratio was found, and effects of changes in energy balance of dairy cows on urinary creatinine excretion should be determined. The urinary excretion of total PD and of allantoin provided lower estimates of duodenal microbial N flow than with measurements in the omasum or duodenum, but they closely reflected the changes observed with these measurements.