Novel fatty acid esters of p-coumaryl alcohol in epicuticular wax of apple fruit

Hexane extracts of epicuticular wax from cv. Gala apples were noted to have an unusual, broad absorbance maximum at approximately 258 nm, which led us to isolate and identify the primary UV-absorbing compounds. Column and thin-layer chromatography yielded a fraction that gave a series of paired, 260-nm-absorbing peaks on C(18) HPLC. These were shown to be a family of phenolic fatty acid esters, for which retention times increased with increasing fatty acid chain length, and paired peaks were esters of two related phenolics with the same fatty acid moiety. Alkaline hydrolysis of the esters released two water-soluble phenolics separable by C(18) HPLC. Electrospray ionization mass spectrometry gave a molecular mass of 150 for both, and (1)H NMR plus UV absorbance spectra identified them as E and Z isomers of p-coumaryl alcohol. Alkaline cleavage of the fatty acid esters in the presence of methanol or ethanol resulted in partial derivatization of E-p-coumaryl alcohol to the corresponding gamma-O-methyl or O-ethyl ether. Gradient HMQC NMR of the HPLC-purified stearate ester of E-p-coumaryl alcohol indicated that fatty acid esterification occurs at the gamma-OH rather than at the 4-OH on the phenyl ring. This is the first report of fatty acid esters of monolignols as a natural plant product.     

Attenuation of Borrelia anserina by serial passage in liquid medium

Borrelia anserina (Sakharoff) was successfully grown in a liquid medium (Barbour-Stoenner-Kelly) for 39 passages. By the 12th serial passage in medium, infectivity of B anserina for chicks was lost. Electron microscopy did not reveal structural differences between non-infective and infective cultured organisms. Changes in the protein profiles were found by electrophoresis as the organisms were passed in culture.     

The intestinal flora of the chicken in the period 2 to 6 weeks of age, with particular reference to the anaerobic bacteria

A study has been made of the intestinal flora of chickens aged between 2 and 6½ weeks. No major differences were found when rations containing 9 or 25% fish meal were compared or when two different sources of fish meal were used. Throughout the period investigated, it was confirmed that the lactobacilli are the only group of organisms generally present in the small intestine in numbers exceeding 104/g. Clostridia were regularly found at 102 to 104/g, but Clostridium welchii was isolated from only occasional samples.

In the caeca, the total number of bacteria present at 2, 3, 4 and 6½ weeks was about 1011/g. Of these it was generally possible to isolate more than 20% using an anaerobic roll‐tube technique and oxygen‐free CO2. Many different types of anaerobes were isolated including strictly anaerobic budding bacteria which had not previously been studied. During the growth of the bird the flora changed. Anaerobic streptococci (peptostreptococci) predominated at 2 weeks but gradually decreased so that by 6½ weeks these organisms formed only a small part of the flora. Many of the Gram‐negative non‐sporing anaerobes only appeared between the 4th and 6th weeks.

Throughout the 2 to 6½‐week period all the caecal samples examined contained 108 to 109 anaerobes/g which were able to attack uric acid and sometimes 1010/g. This property was shared by at least one group of peptostreptococci, four groups of Gram‐negative non‐sporing anaerobes, three groups of Gram‐variable or Gram‐positive anaerobic non‐sporing rods and several species of clostridia.     

Interaction of reovirus and Cryptosporidium baileyi in experimentally infected chickens

No clinical signs, gross lesions, or increased mortality were observed in specific-pathogen-free chickens orally inoculated at 5 days of age with Cryptosporidium baileyi, reovirus 2035, reovirus 2408, or combinations of these agents. Weight gain of chickens inoculated with only reovirus 2408 was depressed 0-8 days postinoculation (PI) (P less than 0.01) but not for the 21-day period PI. Weight gain of chickens inoculated with only reovirus 2035 was not affected. Cryptosporidium baileyi infection significantly depressed weight gain 8-14 days PI but not for the entire 21-day period PI. Weight gain of chickens infected with both C. baileyi and reovirus 2035 was significantly depressed 0-14 days PI and for the entire 21-day period PI. Dual infection with C. baileyi and either reovirus appeared to promote shedding of both agents. Cryptosporidia were found principally in the rectum 2-10 days PI and in the bursa of Fabricius 6-10 days PI. Reovirus infection did not cause any microscopic lesions and did not modify lesions caused by C. baileyi infection.     

Response to selection for milk yield and metabolic challenges in primiparous dairy cows

The effect of selection for milk yield on lactation yield, net energy balance, and on plasma growth hormone, insulin, prolactin, nonesterified fatty acids and glucose was studied in two groups of primiparous Holstein cows of differing genetic merit. Net energy balance was calculated and serial blood samples were collected for a 7 hr period at 0, 45, 90 and 180 days postpartum. Growth hormone releasing factor (.2 μg/kg BW) was administered after 2.5 hr at 0, 45 and 180 days postpartum, while epinephrine (.7 μg/kg BW) was administered at 90 days postpartum, Milk yield was greater, net energy balance was decreased and plasma growth hormone was greater in genetically superior selection group cows compared to control cows.

Growth hormone showed similar increases in both genetic groups in response to growth hormone releasing factor, while prolactin, insulin and glucose were not altered. Epinephrine stimulated an increase in plasma nonesterified fatty acid, glucose and insulin, but responses did not differ between genetic groups. Results indicate differences exist in production efficiency, net energy balance and plasma growth hormone concentration among dairy cattle as a result of selection for milk yield and suggest that selection pressure may act to alter homeorhetic control of nutrient metabolism.     

Vaccination of turkeys with cell-free culture filtrate of Pasteurella multocida: effects of dilution, iron chelation, and heterologous challenge.

Two experiments were done to further define cell-free culture filtrate (CCF) from Pasteurella multocida and its endotoxin content in protecting turkeys against challenge. In the first experiment, the greater-than-30,000-molecular-weight fraction of P. multocida strain R44/6 (serotype 3/4/9/12) CCF was used in 10-fold dilutions given by air-sac inoculation or aerosol to vaccinate turkeys, which were subsequently challenged with either homologous (P-1059, serotype 3) or heterologous (X-73, serotype 1) strains. Endotoxin content of the CCF fraction was high. Compared with positive controls given either live Clemson University vaccine or a commercial bacterin, homologous protection was provided by undiluted CCF and 1:10 dilutions of CCF, but there was no heterologous protection. In the second experiment, CCF of strain R44/6 in regular and iron-limiting media and CCF of strain FC127B (serotype 1/4) were used alone or in combination to vaccinate turkeys, which were challenged as in the first experiment. Homologous but not heterologous protection occurred, even though growth of strain R44/6 in iron-limiting media reduced endotoxin content of CCF by approximately 93%. These results indicate that endotoxin levels of less than 10% but greater than 1% of those in CCF from regular media are sufficient to induce protection in turkeys against homologous challenge but that CCF from either regular or iron-limiting medium does not provide protection against heterologous challenge.     

Nuclear transplantation in bovine embryos

This study was conducted to develop a method for transplanting nuclei in bovine embryos and to test the development of several stages of donor nuclei transplanted to enucleated pronuclear recipient embryos. Pronuclear embryos were centrifuged to reveal nuclei. Nuclei were removed without penetrating the plasma membrane as membrane-bound karyoplasts, and were inserted into enucleated zygotes by electrically induced cell fusion. The highest rate of fusion (79%) occurred in Zimmerman Cell Fusion medium at 100 V for 20 to 40 microseconds with the fusion membranes oriented parallel to the electrodes. The effect of nuclear transplantation on development was tested in pronuclear embryos in which nuclei were removed and reinserted and the embryos were then transferred to sheep oviducts for 5 d. Of the intact nuclear transplant embryos recovered, 5/29 (17%) developed to morulae or blastocysts compared with 11/30 (37%) of the non-manipulated embryos. Two nuclear transplant embryos were transferred to a recipient cow, and both developed to normal offspring. When nuclei from two-, four-, or eight-cell embryos were transplanted to pronuclear recipient embryos, no development was observed.     

Serologic evidence of infectious bursal disease virus infection in Iowa turkeys

Two infectious bursal disease viruses (IBDVs)--a cell-culture-adapted chicken strain designated Edgar strain and a recent isolate from turkeys in Missouri--were used to assay sera from 10 Iowa turkey flocks collected between 1 and 16 weeks of age. The two viruses were serologically distinct in cross-neutralization tests. For all flocks, a similar serologic pattern was found consisting of (1) low maternal antibody titers to turkey IBDV and occasionally to Edgar strain IBDV between 1 and 3 weeks of age, (2) a period of very low or no detectable titers between 3 and 7 weeks of age, and (3) sharply rising high titers to turkey IBDV with low titers to Edgar IBDV beginning at 5 to 8 weeks of age. These findings indicate that infection with IBDV of the serotype represented in this study by the turkey isolate is common in Iowa turkey flocks, whereas infection with IBDV represented by Edgar strain is uncommon. Infection occurred between 3 and 7 weeks of age during the late brooding period or after birds had been moved to an intermediate growing facility. All flocks developed complicated respiratory disease with excessive mortality caused by Escherichia coli septicemia, typically between 3 and 6 weeks of age. Although there was a temporal relationship between IBDV infection and respiratory disease, the possible role of IBDV in the process is unknown.     

Inclusion-body hepatitis in day-old turkeys

Inclusion-body hepatitis was diagnosed in 1-day-old turkeys experiencing above-average mortality. At necropsy, turkeys appeared anemic and had pale yellow livers. Histopathologic examination of affected livers revealed diffuse hepatic degeneration and multifocal necrosis, with approximately 70% of the hepatocytes containing large, basophilic, intranuclear inclusion bodies. An adenovirus was isolated from affected livers and identified as a group I avian adenovirus by indirect immunofluorescence.