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P. E. LAKE

Summary.

Intense acid-phosphatase activity was found in the distal parts of the cells lining the entire length of the vas deferens of the domestic cock and there was evidence that the enzyme(s) is secreted in large amounts into the seminal plasma. Some acid phosphatase(s) was also produced in the vasa efferentia and seminiferous tubules.

Intense alkaline-phosphatase activity was found only in secretions of certain parts of the vasa efferentia, in the tunica intima of all small blood vessels and in intertubular tissue. From the evidence, it is considered that some alkaline phosphatase(s) might possibly diffuse into the seminal plasma from all regions of the genital tract, but it is likely to be small in amount. The significance of the presence of phosphomonoesterases in seminal plasma is discussed.

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P. E. LAKE

Summary.

Fowl semen was diluted 1:3 and stored at 0° to 2° C for 24 and 48 hr before insemination. The diluents were two modifications of a glutamate-containing saline solution calculated to have an osmotic pressure equivalent to 0·98 to 1 % sodium-chloride solution. The concentrations of sodium, potassium, magnesium and glutamate in the diluents were of the same order as those found in seminal plasma uncontaminated with cloacal gland fluid. Calcium was not added and the chloride concentration was low compared with undiluted seminal plasma. One modification of the diluent contained added fructose and the other was fructose-free.

Pullets were inseminated with 0·1 ml of the diluted semen. The proportion of fertile eggs laid by all pullets during days 2 to 6 following insemination with semen stored for 24 and 48 hr in the solution containing fructose was 64 and 47 %, respectively. Fertility was significantly lower in pullets inseminated with semen stored in the fructose-free solution.

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P. E. LAKE and M. HATTON

Semen of the domestic rooster may contain contents of the vasa deferentia with no, or very variable amounts of, `transparent fluid' from structures in the cloaca, depending upon the way in which it is collected by massage (Lake, 1966). The significance of transparent fluid as a component of seminal plasma is not fully understood (Lake, 1966) and thus an examination of all the possible fluid portions of semen, which can be obtained by massage, is important in determining their origin and significance in the reproductive physiology of the fowl. The presence of transparent fluid is detrimental to the prolonged survival of spermatozoa in vitro (see Lake, 1966).

Lake & McIndoe (1959), using paper chromatography, found a large amount of free glutamic acid in the seminal plasma of

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P. E. Lake and O. Ravie

Summary. Improved storage of fowl semen above 0°C was achieved by adjusting the pH of the diluent. The fertility obtained with semen stored for 24 h at 5°C in diluents buffered at different pH values was compared with that of semen stored in a basic, unbuffered solution. The most satisfactory result was achieved with diluent buffered at pH 6·8 or 7·1. Worst fertility was obtained at pH 5·8 and pH 7·4 did not prove very satisfactory. There were indications that the effect of pH under the conditions of the experiment was to regulate metabolism and thereby influence the maintenance of the fertilizing ability of the spermatozoa.

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D. J. BELL and P. E. LAKE

Summary.

A comparison of the phosphomonoesterase activity, against 4-nitrophenyl phosphate, in the seminal plasma of the domestic cock, turkey tom, boar, bull, buck rabbit and of man has been made. The acid-phosphatase(s) activity at pH 4·9 was highest in man, the cock and turkey torn, and lowest in the boar and rabbit, with the bull intermediate. Alkaline-phosphatase activity was highest in the boar and rabbit, lowest in the cock, turkey tom and in man, with the bull again intermediate. It is possible that much of the alkaline phosphatase in the boar semen was derived from the epididymis or testis or both.

Unlike that of the mammals, the acid phosphatase(s) of cock seminal plasma was unable to hydrolyse choline-O-phosphate and the enzyme(s) activity in seminal plasma was only slightly, if at all, inhibited by dextrorotatory tartrate. The significance of these findings has been briefly discussed.

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M. H. EL JACK and P. E. LAKE

Summary.

A method was devised to collect uncontaminated fluid from the uterus (shell gland) of the laying, domestic fowl at two different stages of formation of the egg in the organ.

There was a marked difference between the composition of the uterine fluid found during the early stages of egg formation in the uterus and that found within about 2hr of oviposition. The concentration of sodium fell from 139·1 to 42·8 m-moles/1, whilst that of potassium rose from 15·9 to 75 m-moles/1, at oviposition. The amounts of calcium and magnesium were found to be higher around the time of egg laying and was most probably associated with the deposition of egg shell.

The significance of changes in the ionic composition of uterine fluid to the storage and movement of spermatozoa in the oviduct of the hen has been discussed.

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M. D. TINGARI and P. E. LAKE

Summary.

Ultrastructural studies were made on the uterovaginal sperm-host glands from virgin and inseminated hens. Their fine structure differs only in the presence of spermatozoa in the glandular lumina. Ciliated and non-ciliated epithelial cells were present; the former occurred in the neck region of the tubular glands and merged with the general vaginal surface epithelium.

The true glandular epithelium was composed of non-ciliated cells which showed evidence of high metabolic and secretory activities. The secretion contained carbohydrate and lipid but apocrine secretion rich in glycogen was also observed occasionally. The significance of these secretions in relation to the survival of the stored spermatozoa is discussed. The non-ciliated cells contained many cytoplasmic filaments resembling tonofibrils. It is suggested that these confer contractility on the gland cells, so mobilizing the spermatozoa in response to unknown periodic stimuli associated with oviposition or ovulation.

The stored spermatozoa did not form an intimate association with the lining cells of the glands but the covering membranes of the heads of spermatozoa adhered to each other. It is suggested that this might be due to the absence of antiagglutinating factors in the sperm-host glands.

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A. M. McFARQUHAR and P. E. LAKE

Intra-uterine insemination produces better fertility than intra-vaginal insemination when breeding quail by artificial insemination (Wentworth & Mellen, 1963; Ogasawara & Huang, 1963). Insemination is best accomplished when an egg is present in the uterus. Wentworth & Mellen (1963) passed a needle through the postero-dorsal wall of the cloaca, the uterus wall and the egg, and injected semen into the anterior region of the uterus. Antibiotics were added to the food and to a semen diluent to prevent serious infection of the females by E. coli. Ogasawara & Huang (1963) expelled the egg from the uterus before depositing the semen therein. Good fertility was obtained without the use of antibiotics and infection of hens was not reported.

The present communication reports a simple yet successful method of artificial insemination of quail without expelling the egg from

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W. M. McINDOE and P. E. LAKE

Summary.

The distribution of some hydrolytic enzymes in fowl semen was studied. Washed spermatozoa and seminal plasma were examined for the following enzymes: acid phosphatase, sulphatase, N-acetyl-β-dglucosaminidase, β-glucuronidase, phospholipase A, trypsin-like enzyme and neuraminidase. Only a trypsin-like enzyme, N-acetyl-β-d-glucosaminidase and acid phosphatase were detected in the semen. Most of the acid phosphatase activity occurred in particulate material in the seminal plasma, whereas virtually all the trypsin-like activity was in the spermatozoa. The N-acetyl-β-d-glucosaminidase activity in semen occurred both in the spermatozoa and the seminal plasma; the proportion of the activity in whole semen contributed by the spermatozoa was less than that contributed by the seminal plasma but the concentration of activity was greater in the spermatozoa.

A slightly inhibitory action of seminal plasma on the activity of the trypsin-like enzyme was indicated.

A preliminary examination was made of the semen of different strains of fowl and revealed between-strain differences in the activity of enzymes, particularly acid phosphatase.

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M. D. TINGARI and P. E. LAKE

Summary.

The fate of fowl spermatozoa and testicular fluid retained in the excurrent ducts, and some properties of the epithelial cells lining the ducts, were studied after ligation of the ductus deferens by light and electron microscopy. Striking changes occurred 3 or 4 weeks after ligation.

Spermatozoa accumulated cranial to the ligature chiefly in the lumina of the ductuli efferentes. They showed signs of disintegration mainly in the head where there was a disruption of chromatin and loosening of membranes. An increased amount of cell débris of unknown identity was also observed in the lumina of the ducts.

The uptake of spermatozoa by epithelial cells lining the male tract was seen in normal males but was much increased after ligation. The process was evident in the low cuboidal cells of the rete testis and in the ciliated and non-ciliated Type I cells lining the ductuli efferentes and narrow connecting ductules, and in the non-ciliated Type II cells lining the wide connecting ductules, ductus epididymidis and ductus deferens. Macrophages, containing spermatozoa, were found in the lumina of the ducts, in the subepithelial tissue and wedged between the basal lamina and the surface epithelia. All of these observations may indicate a route for the disposal of unejaculated spermatozoa in the male fowl.

Cytoplasmic vacuoles containing testicular fluid were seen in the apical parts of the ciliated cells and this may represent resorption of the fluid by these cells.