summary of the phd thesis · of puerperium over the reproductional activities in two farms (farm a...

Managementul puerperiumului la vacă şi importanţa lui în reproducţie

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UNIVERSITY OF AGRICULTURAL SCIENCES AND VETERINARY MEDICINE

CLUJ-NAPOCA PHD SCHOOL

FACULTY OF VETERINARY MEDICINE

EERRCCEEAANN EELLEENNAA

SUMMARY OF THE PhD THESIS

COW POST‐PARTUM MANAGEMENT AND ITS IMPORTANCE IN REPRODUCTION

Scientific coordinator:

Prof.univ.Dr.Claudius Lisovschi‐CHELEŞANU


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RESEARCH PURPOSES

Milking cows breeding and reproduction in private exploitations of familial or

associative type has an enormous importance being very actual and with high

expectances in the future.

Continuous improvement of human food industry nationally and worldwide

requires a high production of milk and meat at minimal costs on product unit, that can be

obtained by elevating the productive performances and animal genetic amelioration.

Considering the fact that milking cows exploitation is based on the knowledge of the

animal’s individual particularities and the connections that are revealed between the

genotype and the environment, for obtaining a optimal production for the genetic

potential, we need to know the exploitation factors, the organization of production-

reproduction, animals health status and the exploitation management.

The thesis research purposes are to highlight the importance, evolution and impact

of puerperium over the reproductional activities in two farms (Farm A and Farm B) and

his implications over the economics and profitability of milking cow farms.

The established objectives for the purposes mentioned were the following:

evaluation of the natural and organisational environment in which the

researches were carried

clinical evaluations regarding puerperal diseases in milking cows and

their implications in the service-period evolution

laboratory evaluations regarding milking cows puerperal diseases

ultrasound aspects in milking cows puerperium

puerperium influence over milking cows reproduction and his zoo-

economical implications

the therapeutical conduct in puerperal and post-puerperal diseases of

milking cows


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CHAPTER IV THE NATURAL ENVIRONMENT AND ORGANIZATIONAL SETTING IN

WICH THE RESEARCHES WERE CONDUCTED

Researches were conducted during 2005-2010, having the organizational setting for the proposed objectives two cattle farms in Mures County. We observed two bovine populations from two breeding and milk production exploitation farms, respecting the general conditions of microclimate, alimentation and maintenance.

The first farm (Farm A) has an intensive husbandry system, cows being tied on two lines – head to head – separated by an alley. The total bovine effective was of 290 heads from which 62 were in lactation. The second farm (Farm B) has a free husbandry system, with untied animals, permanent stabulation, with spaces for rest and feeding. The total effective of farm B was 300 heads, from which 68 were in lactation. The research material was constituted by cows from the following breeds: Holstein-Friesian, Romanian Spotted cow, Frisona and Red Holstein.

Fig. no. 4.1. Aspects from Farm A maintenance system

The milking was carried out twice a day with a mobile milking device (two for

every farm). The cow maintenance system from farm A during fall-winter period was represented by permanent stabulation, with tied animals. On spring-summer period, the cows were let free on the pasture, this being provided with ground paddock with shade and space for movements. At farm B, the maintenance system the same for all year, with space for movements, feeding space, rest beds, the animals being untied all the time. In the last period of gestation the cows were moved in individual calving spaces.


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Fig. no. 4.2. Aspects from Farm B maintenance systems The feeding was made with the main forage resources existent in the area,

considering their nutritive value and the daily quantity that a cow can eat in a day. Feed rations were of a good quality, the feeding being done according to the season, physiological state of the animal and the milk production. Summer feed rations were characterized by: lowering the corn silo, adding green alfalfa at the total basic ration, approximately 30 kg a day. Detailed knowing of bovine categories, conformational development, morpho-physiologic aspects and behavior, constitute extremely useful basic elements for establishing the technological fluxes for well determined economic purposes.

CHAPTER V

CLINICAL RESEARCHES REGARDING PUERPERAL PATHOLOGY IN MILKING COWS AND THEIR IMPLICATIONS IN SERVICE-PERIOD

EVOLUTION

The objectives of the study were represented by monitoring before and after birth of milking cows especially the puerperal period, mainly focusing on the following parameters: cow welfare, alimentation, environment, the animal health state and the microclimate with their implications in the service-period evolution until the installation of a new gestation.


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The aim of this study was accomplished by making two experiments. In the first one we evaluated 10% of the selected cows and kept them under supervision until day 120 after-birth, tracking the birth-insemination periods for a overview of the reproductive activity in both farms. The second experiment had the purpose of monitoring the puerperium evolution and the incidence of after-birth pathology at the entire heard of both farms.

5.2 MATERIAL AND METHOD

For the first experiment researches were made during 2009-2011, in two milking

cow farms, of Holstein-Friesian, Romanian Spotted cow, Frisona and Red Holstein breeds (10 from each farm). The exploitation conditions were relatively good, with variable feeding according to the physiological state.

Three weeks before the parturition, the cows got a excess of concentrated feed. The cows selected for the study had ages between 3 and 7 years, being constantly watched for three weeks before parturition and 120 days after that. The scoring was carried out along with a specialist, and the grades being between 0 and 5.

For the second experiment three batches were established in each farm: Batch M, the witness batch constituted from cows with normal puerperium, batch E1 constituted from cows with puerperal pathology and batch E2 constituted by cows with ovarian pathology. Batches were constituted from cows in different periods from parturition, being examined clinically according the gynecological enquiry. Cows were clinically observed in general (case history, inspection, palpation, temperature) and by special gynecological exams made by trans-rectal palpation, and colposcopy.

The trans-rectal examination established the consistency changes, volume, uterine tonus and the aspects of uterine secretions, as well as the ovaries regarding their shape, volume and surface or depth formations.

5.3 RESULTS AND DISCUSSIONS

In the first experiment, cow ratings revealed that 80% of the animals from farm A

were rated with 3 and for the farm B only 60%, revealing worst general state for the animals in farm B.

Table no. 5.1. Cow ratings results for both farms

No.

Corporal evaluation / indicative

Number of cows Percent %

Farm A Farm B Farm A Farm B 1. =3 8 6 80 60 2. <3 2 4 20 40

In farm A no cow in heats was identified and no insemination was made in the first 40 days after parturition. In farm B was identified and inseminated only one cow in heats. In the 40-70 days period after parturition, in farm A were identified and inseminated seven (70%) cows in heats and in farm B were identified and inseminated


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only four (40%) cows in heats. For the period of more than 70 days after parturition, in farm a were identified and inseminated 3 cows (30%) and in farm B 5 cows (50%).

01234567

First period under 40 days

Second period 40 ‐70 days

Third period , more than 70 days

0

7

3

Number of cows

Chart no. 5.1. Cows in service-period and heats revealing period

for cows in farm A

0

5

1 period less than 40 days

2 period 40‐70 days

3 period, more than 70 days

1

45Number of cows

Nr

Chart no. 5.2. Cows in service-period and heats revealing period

for cows in farm A

In farm A, from the 10 cows taken in the study, 9 were diagnosed with pregnancies (7 in the 40-70 days period and 2 in the 70-110 days period), only one wasn’t pregnant and was removed from the heard. In farm B, from the 10 cows of the study, two weren’t diagnosed pregnant (later being taken to the slaughterhouse, five were diagnosed with pregnancy in the 40-70 days period and three cows were diagnosed with pregnancy in the 70-110 days period (for a total of 8 cows with pregnancies in both periods). The farm A had three (30%) cows that repeated the heats after the first artificial insemination, and farm B had five (50%) cows with the same problem. After the second artificial insemination, in farm A only one cow repeated the heats (10%) and in farm B were three cows (30%) that repeated the heats.

After the third artificial insemination all cows got pregnant in farm A and two repeated the heats in farm B (20%).


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Chart no. 5.3. Heats repeating frequency after inseminations at farm A and B In the second experiment of this chapter, the following aspects were revealed: In many cases the exteriorized uterine horn was dirty, covered by fetal annexes,

with different lesions from ruptured mucosa till uterine artery rupture, caruncles rupture and death of the animal. The main puerperal problems clinically diagnosed were fetal annexes retention, prolapsed uterus, local and general puerperal infections, atone uterus afterbirth, ovarian diseases. The most common puerperal problems found in both farms were puerperal infections (on every level of the genital area) - usually found after fetal annexes retention, prolapsed uterus afterbirth – and the ovarian pathology. The researches reveal an increased rate of puerperal affections during periparturial morbidity. The incidence of puerperal affections was demonstrated after research on 207 animals of local breeds (table no. 5.2.).

Table no. 5.2 Puerperal diseases incidence for cows from farm A and B

Number of cows

Uterine atony post-

partum

Fetal annexes retention

Uterine infections

Ovariopaties

Total Total

Corp luteal Luteal body

Hypotrophic

ovaries

Ovarian

cysts

N % N % n % n % N % N % N %

207 20 10,2 20 10,2 38 19,3 40 20,3 26 13,2 8 4,1 6 3,0

4

The incidence analysis shows that the uterine atony had a 10,2% value, fetal annexes retention and genital infections had 19,3% and ovarian diseases had a 20,3% incidence. 41,5% of the cases had puerperal affections after periparturial morbidity. The parturition-insemination periods, service period and calving interval showed different values according to the diagnosed pathology.


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Table no. 5.3 Average values of PI, SP and CI periods at cow from both farms

Number

Intervals

Births

Batch M Batch E1 Batch E2 Cows with normal

puerperium Cows with

puerperal diseases Cows with ovarian

pathology 1. No cows days No cows days No cows days 2. P-I 207 109 70 ± 2 58 122 ± 5 40 120± 5 3. SP 207 109 94± 4 58 145± 8 40 170±10 4. CI 207 109 371± 3 58 420± 8 40 445±10

Legend: P-I= parturition-insemination interval , SP= service period; CI= calving interval

About the influence of puerperal pathology over the reproduction parameters, the average parturition-insemination period was of 70 ± 2 days for the normal cow puerperium (witness) batch M, of 122 ± 5 days for the cows with puerperal affections and 120 ± 5 days for the cows with ovarian pathology. The service period had bigger values for the females with affections compared with the witness batch, the same aspects were found for the calving interval.

CHAPTER VI PARACLINICAL RESEARCHES REGARDING PUERPERAL AFFECTIONS IN

COWS

6.1. RESEARCH PURPOSES

Bovine management asks for interdependence between the following factors: nutrition, production and reproduction.

The progresses obtained in bovine genetic amelioration had for an effect the obtaining of two types of cows with productive performances, but more sensitive to different pathogens or conditioned-pathogens because of an organic and homeostatic balance labile for the smallest perturbations of feeding factors, hygiene factors or media factors.

A permanent evaluation of cow health is imposed, by establishing the hematologic and biochemical profile for the entire heard.

The objectives were constituted by: establishing the hematologic and biochemical profile of the animals, microbiological determinations, macroscopic and microscopic aspects, ultrasonographic aspects of cow puerperium, considerations regarding the puerperium influence on milking cows’ reproduction and their zoo-economical implications.


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6.2. HEMATOLOGIC AND BIOCHEMICAL PROFILE DETERMINATION

6.2.1. Material and method Researches were realized in both professional exploitations (A and B), by making

two batches of animals for every farm: witness batch M1 – formed by cows with normal puerperium, experimental batch E1 – composed from cows with puerperal pathology, witness batch M2 – formed by cows with normal puerperium and batch E2 – constituted from cows with ovarian pathology. Every batch had 10 females with variable puerperium (between 0 and 110 days after parturition).

Hematological determinations were realized using a automatic analysis device MS 9-5 Mellet Schloesing, that evaluates 29 parameters, from witch: erythrocytes, blood plackets, leucocytes, from blood harvested with EDTA. For serum proteins, protein fractions (albumins, globulins), cholesterol, blood sugar, enzymatic profile, mineral profile, KY190 (260 tests per hour) biochemistry-turbidimetry automatic analysis device was used by colorimetric method in continuous flux.

6.2.2. Results and discussions Hematological examinations results for the studied cows relevated different

variations for the main hematological parameters for cows with reproduction problems compared with healthy cows and the scientific literature. For the cows with puerperal affections (batch E1) erythrocytes values were smaller than the ones of the M1 batch. The erythrocyte number (mil/mm3) had an average of 5,65 ± 0,30 at E1 batch, smaller than the M1 batch number 6,30 ± 0,79 mil/mm3. The difference was insignificant being situated at the minimal limit of the references quote.

The haematocrit had variations around 23,80 ±1,55 for batch E1 and 33,50 ± 0,30 for batch M1, the difference is significant. Hemoglobin (Hb, g/dl) had an average value of 8,5 ± 0,50 for E1 batch and 11,30 ± 0,25 in batch M1. The difference between the two batches was significant, this anemia being explained by the high productivity and the extremely high exploitation of the studied cows. VEM (µ³) and HEM (pg) presented values very close to the maximum limit of reference quote, both for batch E1 and M1, explained by the body tendency of anemia compensation by releasing big erythrocytes in the blood and the increasing productivity. CHEM (g/dl) had values found in the middle line of references quotes (26 - 34 g/dl), with an average value of 30,90 ± 1,10 for batch E1 and 32,50 ± 2,30 for batch M1. The value difference is insignificant. Total leucocytes number had an average value of 13,30 ± 1,10 x 106 /mm³ compared to the witness batch M1 7,75 ± 1,00 x 106 /mm³, a distinctly significant difference. Leucocytes line elements had average values of lymphocytes in batch E1 insignificantly smaller than batch M1, the same levels were found in monocytes but neutrophils, eosinophils and basophiles had higher values with significant differences between batches E1 and M1 and reference quote.


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For the cows from batch E2 with ovarian pathology erythrocytes constants had different variations from the witness batch M2 and reference quote. Erythrocytes had average value of 5,80 ± 30 in batch E2 comparing to batch M2 5,70 ± 0,91, insignificant value.

Hematocrit had average value of 26,35 ± 0,50 for batch E2, in compare with batch M2 = 32,25 ± 0,30. Hemoglobin was around 8,80 ± 0,40 in batch E2 instead of 12,60 ± 0,20 in batch M2.Both hemoglobin and hematocrit showed values around the minimal limit of reference quote in batch E2 (hemoglobin had value under the lower limit of reference quote 8 – 15 g/dl compared to witness batch M2, in which the values were at the higher limit of reference quote. These low values of hemoglobin and hematocrit show a variable anemia for the studied cows. VEM and HEM registered average values situated at the higher limit of reference quote for both batches, E2 and M2. Like in the previous experiment this can be explained by an anemia compensation tendency of the body. Total leucocytes number registered a average value of 9,45 ± 0,70 in batch E2 instead of 7,26 ± 1,04 at batch M2, founding a significant difference between the two batches. Lymphocytes and neutrofils presented average values, situated at the highest limit of the reference quote, the difference between the two batches being insignificant. Eosinophils and monocytes showed values around the highest limit of reference quote for batch E2, the difference between these values and the ones of batch M2 being significant. Leucocytes formula results for batches E2 and M2 show the presence of inflammations and ovarian dysfunctions. The protein profile determination was constituted from total serum proteins determination and urea determination from blood serum. Total serum proteins (g/dl) registered in batch E1 (the one with puerperal diseases) had an average value of 6,30 ± 0,7 compared to the witness batch value 9,10 ± 0,50. In batch E2 (composed by cows with ovarian pathology) the average value was of 6,80 ± 0,65, and in the witness batch from the same farm (M2), the average was 8,75 ± 0,10.

The values obtained are considered to be in the physiological limits for bovines, with a small amendment, that experimental batches E1 and E2 had average values of total serum proteins much lower than witness batches and the reference (7,00 – 8,05), indicating a disequilibrium in the protein metabolism. Urea values for batch E1 had average values of 33,00 ± 0,30 comparing to the witness. In batch E2 the average values were 14,80 comparative to 18,35 in batch M2.

The results show that for batches E1 and M1 urea had an average value situated in the higher limit of reference quotes (20 – 40 mg/dl), explained by a food ratio rich in nitrous substances without organic dysfunctions in the studied animals. For batches E2 and M2, average values of urea (14,8 respectively 19,3) are situated in the lowest limit of reference quote, resulting low in nitrate intake. We can conclude that we can speak of a disorder of the protein metabolism in all the four batches, reason for which we recommend a equilibrated ratio.

The energetic profile was composed from determination of glycemia and cholesterol. Glycemia showed a average value of 51,60 ± 0,50 in batch E1 and 58,30 in the witness batch M1, without significant differences for batch E2 average value was considered to be 75,80 compared to witness M2 with 77,30. Glycemia results show values situated at the low limit of reference quote for batches E1 and M1 (green grass –


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hypoglycemia) and for batches E2 and M2 the values were at the superior limit of reference quote. Cholesterol registered in batch E2 a average value of 205 ± 8,50 instead of witness batch M2 with an average value of 196 ± 5,10. For batch E1 the value was 218 ± 15,10 comparing to M1with an average value 220 ± 5,20

In batches E1 and M1 we considered a hypercholesterolemia (plus a hypoglycemia) because the lack of green fodder in the ratio. For batches E2 and M2 the cholesterol average values were situated in the superior limit of reference quote (100 – 200 mg/dl).

The mineral profile was composed by serum determinations of calcium, phosphorus and magnesium. Blood calcium (8-11 mg/dl) showed an average value of 8,70 ± 0,30 for batch E1 in compare with batch M1 11,22 ± 2,10. Batch E2 showed an average value of 9,30 ± 1,20, and M2, 10,40 ± 2,05. Phosphorus (5,5-10 mg/dl) had an average value of 6,35 ± 2,10, in batch E1 and batch M1 8,90 ± 2,30. Batches E2 and M2 had different values also 4,90 ± 1,50, respectively 6,80 ± 2,30.

Magnesium (1,50 - 2,50 mg/dl) showed an average level of 2,06 ± 0,15, for batch E1 and 2,36 ± 0,20, for batch M1. Batch E2 had an average value of 2,53 ± 2,12, and the witness M2 batch, 2,82 ± 0,75. The phosphorus/calcium ratio is modified because of the lowest limit of reference quotes in calcium and phosphorus for batches E1 and E2 compared to the witness batch that had average values of phosphorus and calcium found in the superior limits of the reference quote.

The magnesium levels showed that batches E1 had levels situated in the inferior level limit of reference quote compared to witness M1. Batches E2 and M2 had values situated at the superior limit of reference quote.

The serum values difference of calcium, phosphorus and magnesium in the two experiments were insignificant. The registered results showed some metabolic deficiencies at cows with puerperal affections and at those with ovarian pathology, a fact that imposes as a major request establishing the metabolic profile for cows as a permanent health surveillance test.


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Table no.6.1. Hematological results obtained in the E1 and M1 batches of cows

No.

Parameters

Measure

Unit

Normal values

Batches

E1 M1

Variation limits Variation limits

1. Erytrocites(E) mil/mm3 5-10 5,65 0,30 5,20-5,80 6,30 0,79 5,90-7,10

2. Hematocrit (Ht) % 24-46 23,80 1,55 24,50-30,70 33,50 0,30 31,70-36,60

3. Hemoglobin (Hb) g/dl 8-15 8.05* 0,50 7,20-10,10 11,30 0,25 9,20-14,90

4. VEM 40-60 59,30 6,30 48,80-60,30 57,30 5,30 43,80-60,00

5. HEM pg 11-18 22,90 2,30 12,20-24,10 22,20 3,20 19,50-24,00

6. CHEM g/dl 26-34 30,90 1,10 28,90-33,80 32,50 2,30 34,00-40,00

7. Total leucocyte number X 106./

4-12 13,30** 2,10 10,20-14,80 7,75 1,00 7,60-8,10

8. Limfocytes % 40-60 49,90 1,50 45,80-54,60 54,00 2,50 50,70-60,40

9. Neutrofils % 15-47 55,75* 0,60 52,50-60,70 35,90 1,60 31,80-38,10

10. Eozinofils % 2-10 6,80* 0,55 3,90-10,70 2,90 1,66 2,20-3,60

11. Basophils % 0-4 1,77* 0,30 1,20-2,60 0,50 0,30 0,90-1,40

12. Monocites % 2-6 3,10 1,30 2,40-5,10 3,30 2,50 1,60-2,90

- average, - standard variation, *p<0,05 –semnificative difence, ** p=0,1 –distinct semnificative diference


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Table no.6.2. Hematological results obtained in the E2 and M2 batches of cows

No.

Parameters

Measurement

unit

Normal values

Cow batches

E2 M2

Variation limits Variation limits

1. Erytrocites(E) mil/mm3 5-10 5,80 0,30 5,90-6,90 5,70 0,91 5,30-5,60

2. Hematocrit (Ht) % 24-46 26,35 0,50 26,00-34,00 32,25 0,30 30,50-35,00

3. Hemoglobin (Hb) g/dl 8-15 8,80 0,40 8,40-10,80 12,60 0,20 9,40-13,80

4. VEM 40-60 50,48 3,60 43,00-51,00 57,50 4,50 45,10-58,90

5. HEM Pg 11-18 16,95 2,80 13,70-18,10 21,90 3,20 19,70-24

6. CHEM g/dl 26-34 32,06 2,40 30,90-34,00 35,90 2,30 33,60-49,00

7. Total leucocyte number mii./ 4-12 9,45* 0,70 8,70-11,60 7,26 1,04 7,10-7,30

8. Limfocytes % 40-60 54,75 3.05 50,05-58,00 58,80 2,80 56,20-60,00

9. Neutrofils % 15-47 40,75 2,20 35,00-42,00 38,66 1,60 33,00-40,00

10. Eozinofils % 2-10 4,50* 0,03 4,30-4,80 2 1 1,80-2,20

11. Basophils % 0-4 2,45 0,03 2,00-2,75 1,76 0,33 1,00-2,40

12. Monocites % 2-6 4,25* 0,03 2,00-6,45 2,10 1,00 1,40-3,00

- average, - standard variation, *p<0,05 –semnificative difence, ** p=0,1 –distinct semnificative diference


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6.3. MICROBIOLOGICAL ASSAYS

The bacterial infections from different segments of the cow genitalia constitute one of the most frequent problems found in the veterinarian practice. The infections evolution may vary a lot being dependent of multiple factors. Bacteria identification was possible after isolation in pure culture from the tested sample followed by the study of their morpho-biological characteristics. The bacteriological examination had in generally the following steps: sample harvesting, direct examination from the pathological material, examinations from cultures and bacteria identification by their characteristics.

6.3.1. Material and method The bacteriological examination was performed on samples gathered from 20

cows in the first four weeks after parturition, from both farms A and B, with different pathologies. For the tests we harvested genital secretions in sterile recipients from the different levels: vagina, vulvae and uterus. Smears and cultures were made with different culture media and in different growing conditions for bacteria species identification.

The identifications of isolated germs was made according to the cultural, bacteriological and biochemical examinations. Germ sensitivity was tested using the difusometry method, using the antibiotics micro-tablets and reading the inhibition areas diameters.

6.3.2. Results and discussions The bacteriological analyses showed bacteria in unique culture, in mixed culture

and in association culture. For the cows with pathologic puerperium the most frequently were isolated bacteria from the following species: Arcanobacterium pyogenes, Escherichia coli, Streptococcus and Staphylococcus.

Fig. no. 6.1.Mixed culture of staphilococcus and streptococcus

Fig. no 6.2 – Mixed culture of staphylococcus and arcanobacterium.


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After the evaluations, the following aspects were shown: Total germ number grows progressively until maximum values in the first two weeks after-birth, with significantly higher levels at cows with dystocic labours, with the predomination of isolated bacteria in pure culture (for Escherichia coli, TGN 106= 250, Arcanobacterium pyogenes TGN 106= 236) and in mixed culture (Actinomices bovis (120) şi Proteus spp (120), Arcanobacterium pyogenes (215) şi Escherichia coli (215), Arcanobacterium pyogenes (58,6) and Streptococcus spp (45 – 72);

Isolated bacteria species were Gram positive in 52% of cases and Gram negative in 48%. For the cows with dystocic parturitions mainly (50%) were bacteria from associated cultures. In the 4th week after birth were isolated germs only from the dystocic parturitions, from which 50% were constituted by Arcanobacterium pyogenes.

The main bacteria species found were: 33,3% Escherichia coli (Gram negative) in mixed culture, 50% Arcanobacterium pyogenes (Gram positive) in mixed culture, Staphylococcus (Gram positive), 16,6% Actinomices bovis (Gram positive) in pure and mixed culture, 16,6% Streptococcus spp. (Gram positive) in pure and mixed culture, 16,6% Proteus spp. (Gram negative) in mixed culture, 16,6% Bacillus cereus (Gram positive) in pure culture, 10,5% Bacteroides spp. (Gram negative) and Fusobacterium necrophorum (Gram negative) in association culture.

The bacterial incidence in the genital secretions shows that in the examined samples is a balance between the species with affinity for Gram coloration. The germ sensibility for the used antibiotics was made by measuring the inhibition diameters, with lack of germ colonies, as the surface is more extended, the antibiotic is more efficient.

According to antibiogram (figure 6.5.) the germs were sensitive for Flumequine (inhibition area of 30 mm) and Enrofloxacin (inhibition area of 26 mm); the germs were moderately sensitive to Ampicilin (inhibition area of 20 mm) and resistant to Eritromicin (without inhibition area).

Pseudomonas, Escherichia coli and Proteus (Fam. Enterobacteriaceae) were sensitive to: Amoxiklav (inhibition area of 18 mm), Gentamicin (inhibition area of 17 mm), Doxiciclină (inhibition area of 16 mm). They were resistant to Ampicilin (inhibition area of 10 mm) and Eritromicin (inhibition area of 11 mm). Staphilococcs were sensitive to: Penicilin (inhibition area of 29mm), Amoxiclav (inhibition area of 22 mm), Kanamicin (inhibition area of 18 mm) şi Flumexin (inhibition area of 30 mm). They were resistant to Erythromycin (inhibition area of 4 mm) şi Ampicilin (without inhibition area). Streptococcs and Arcanobacterium were sensitive to: Eritromicin (inhibition area of 22 mm), Tetracilcin (inhibition area of 25 mm) and they were resistant to Gentamicin, Ampicilin, Penicilin (without inhibition areas). Most samples showed bacteria specific and unspecific for bovine genitalia.


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Fig.no. 6.5 – Mixed culture antibiogram

6.4 HISTOLOGICAL ASPECTS

6.4.1. Material and method Investigations were made on genital organs from 20 cows from the breeds showed

before, that were slaughtered during 2007-2010 from farms A and B. Fragments of uterus, uterine horns, cervix, ovaries and saplings from cows with different genital pathologies, were harvested and prepared in the UMF Tg. Mureş Laboratory.

The tissue and organ fixation was made immediately after harvesting in formaldehyde 10%, the colorations used were HE (hematoxylin-eosin) and HEA (hematoxiline – eosin – methyl blue) for tissue main aspects and Tricrom Masson, MGG (May Grunwald-Giemsa) for the cellular details.

6.4.2. Results and discussions Examination results from cows with genital puerperal pathology showed

macroscopical and microscopical aspects (histopatological) specific to puerperal diagnosed pathology: vulvae-vaginal affections, uterine affections, ovarian pathology.

C. Macroscopical aspects The macroscopic examination of 5 slaughtered cows showed: infiltration in vulvae

labia, agglutinated hairs, vaginal mucosa of purple color with erosion areas, crushed tissues covered with fake membranes, abscesses and fistulae.

The uterine lesions present in the genital pathology are frequent in all the uterine segments being showed in 10 cows from the 20 tested. Cervical lesions are macroscopical characterized by: bolded folds, paste, spongious consistency and muco-purulent collection accumulated in the vaginal ends.

In the tested cows, the cervix pathology was associated with metritis and vaginitis, having a ascendant or descendant evolution 9from the vagina or the uterus).

The persistent lutheal body was showed at 5 cows from the 20 slaughtered.


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D. Microscopical lesions

In the pathological material harvested from the cows with puerperal vulvae-vestibule-vaginal pathology, we found macrophages in phagocytosis process, and in inactive state.

Fig. no. 6.6. Macrophages in phagocytosis process

At the cervical level, histopathological nuclear hyperchromatosis was showed in

the cervical musculae.

Fig. no. 6.7. Nuclear hiperchromatosis

In chronic endometritis limpho-granulocitary infiltrate was obvious, peri and

intraglandullar difuse and in centered fig. no. 6.7) or microabcesses in the uterine stroma (fig.no.6.8) and a active phagocytosis process made by the periglandular macrophages.

Fig. no. 6.7.

Centered limpho-granulocitary infiltrate peri and intraglandulary

Fig. no. 6.8. Microabcesses in the uterine stroma


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In the starting phase of chronic endometritis chistical hyperplasia was obvious along with glandulo-chystic degeneration with diffuse lympho-histiocitary infiltrate and fibrosis tendency (fig. nr.6.9) and glandular hypoplasia with massive fibrosis without reticulohistiocitary reaction (fig. nr.6.10).

Fig. no. 6.9. Glandulo-chystic degeneration

Fig. no. 6.10.Endometriosis with glandular hypoplasia

In the cases of luteal body persistence on the ovary associated with chronic

endometritis, histological examination revealed necrobiosis, vacolar degeneration and blastic hyperplasia. Also limpho-histiocitary infiltration was observed with macrophages histiocytes in different activity stages and chystic uterine glands. In the chronic forms chistical hyperplasia development was observed, with mucus and neutrophile granulocytes in different stages of cariopicnosis, cariolysis and cells from the limpho-granulocitary infiltrate in different necrobiosis stages.

6.5. ULTRASONOGRAPHY ASPECTS IN COWS PUERPERIUM

6.6.1. Material and method

Researches were made on 20 cows, 10 from each farm (2 farms), for the purpose

of finding by using the ultrasonography device along with the clinical exam the following physiological aspects: uterine volume reduction, number of blood vessels and their caliber, quantity and quality of uterine content, ovarian activity starting in correlation with the appearance of follicles in different development stages.

For these purposes we used a portable ultrasonograph device (Figura nr.6.11) Esoate Aquila Pro with trans-rectal linear probe at a frequency 5 MHz.

Fig. nr. 6.11. Portable ultrasonograph device used in the study

Fig. nr. 6.12. Cow uterus at 17 days postpartum


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6.5.2. Results and discutions The ultrasonograph examination of cow ovaries and uterus during the puerperal

period showed different physiological and pathological aspects by a variable imagistics according to the diagnosis. From the total of 20 cows tested the followings were found: - In farm A from the total of 10 cows, three (30%) presented pathological puerperium, two of them (20%) being diagnosed with lochiometritis and one (10%) with purulent metritis. - Seven of the cows (70%) showed physiological puerperium, with a normal involution of genital segments and follicular development characteristic of the period. - In farm B, from the 10 cows studied, four (40%) presented pathological puerperium, characterized by lochiometritis 2 cows (20%) and purulent endometritis 2 cows (20%). The animals in physiological puerperium were 60%.

6.6. PUERPERIUM INFLUENCE OVER MILKING COWS REPRODUCTION CAPACITIES AND THEIR ZOO-ECONOMICAL IMPLICATIONS

The development of all biological processes ensures the organism adaptation on

different request, including the organism production and reproduction necessities. In metabolic and nutritional imbalance, the cow’s heath status is affected in

general and especially their reproductive capacities, causing reproductive, productive and economical parameters decrease.

6.6.1. Material and method Researches were carried out during 2007-2010, on cows and heifers from the two

farms studied, with ages between 3 and 7 years, in variable gestational and lactation periods. Cows were examined by using the gynecological enquiry, being monitored using more criteria: artificial examination index, fecundity, birth rate, calving interval and service-period (SP).

6.6.2. Results and discussions The reproductive attributes like birth-rate (N), fecundity (F), calving interval (CI),

and service period (SP) have direct influence over the herd rhythmical growth, with important zoo-economical implications.

In the year 2007, in farm A, which had an effective of 170 cows and heifers, 110 have given birth, 58 were artificially inseminated and 49 got pregnant. In farm B, from a herd of 190 cows and heifers, 126 have given birth, 69 got artificially inseminated and 50 of them got pregnant. According to the number of inseminations, in farm A 7 cows got inseminated 4 times, 2 being reformed and slaughtered and in farm B 10 cows got inseminated 3 times, seven got inseminated 4 times and 2 cows were reformed and slaughtered.


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In the year 2008, in farm A, which had an effective of 180 cows and heifers, 131

have given birth, 73 were artificially inseminated and 64 got pregnant. In farm B, from a herd of 201 cows and heifers, 129 have given birth, 75 got artificially inseminated and 55 of them got pregnant. According to the number of inseminations, in farm A 11 had repeated inseminations because of the uterine infections and in farm B 10 cows got inseminated 3 times, six got inseminated 4 times and 4 cows were reformed and slaughtered.

The year 2009 had the following data regarding the two farms: in farm A, which had an effective of 186 cows and heifers, 143 have given birth, 88 were artificially inseminated and 75 got pregnant. In farm B, from a herd of 208 cows and heifers, 130 have given birth, 77 got artificially inseminated and 53 of them got pregnant. According to the number of inseminations, in farm A 13 cows got inseminated multiple times until they got pregnant, and in farm B 11 cows got inseminated 3 times, seven got inseminated 4 times, 3 cows were reformed and slaughtered and 3 cows had an unsure situation, being proposed for slaughtering.

In the year 2010, in farm A, which had an effective of 196 cows and heifers, 148 have given birth, 92 were artificially inseminated and 81 got pregnant. In farm B, from a herd of 220 cows and heifers, 133 have given birth, 72 got artificially inseminated and 57 of them got pregnant. According to the number of inseminations, in farm A 9 cows received repeated inseminations because of the uterine infections, two of them got hysterectomy. In farm B 3 cows got inseminated 3 times, seven got inseminated 4 times and 2 cows got inseminated 5 times.

Table no.6.6 2007 – 2010 farm A and B statistics for necesity sacrifications and reformations

No. Criteria

Farm A Farm B Year Year

2007 2008 2009 2010 2007 2008 2009 2010

1. Genital problems 2 - - 1 2 4 3 1

2. Medical problems 2 - 3 1 - 1 4 3

3. Mammary gland problems - 2 - 1 1 - 3 -

4. Leg problems - 2 3 2 - 2 1 2 5. TOTAL 4 4 6 5 3 7 11 6

Fecundity index in insignificantly larger in farm A than farm B, because the artificial insemination index and service period were smaller. For both farms a economical efficiency growth is necessary by monitoring with particular attention the puerperal period by improving the sanitary-veterinary activities, improvement of all technological parameters, increasing the production level, superior qualitative application of production and cost reduction on product unit.


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CHAPTER VII

THERAPEUTIC CONDUCT IN PUERPERAL AND POSTPUERPERAL DISEASSES IN MILKING COWS

The established objectives were:

• implementation of a therapy for chronic puerperal endometritis treatment specific for the tested territory (experiment 1); • implementation of a therapy for reproduction disorders determined by the luteal body (experiement 2); • determining a adequate therapy for estrus and ovulation induction and synchronization for the studied milking cows (experiment 3).

MATERIAL AND METHOD Researches were made on 60 dairy cows, form the following breeds: Romanian

spotted cow, Red Holsthein, Holsthein Friesian and Frisona, with ages between 3 and 7 years, with different pathological genital diseases. We used associated and unique therapy. Three experiments were made during two years (2008-2010):

experiment 1 – was made on three batches of 10 cows each, diagnosed with

puerperal endometritis (8 cows) complicated with pyometra (2 cows). The cows from batch E1.1. were treated with the product Estrumate® (500 mcg/dose) and after 2-4 days with the product Metricure® ( one syringe intrauterine) , the cows from batch E1.2. were treated with the product Metricurine, (homeopath drug), 10 ml/dose/animal, from two in two days, during 6 days. The cows form the whitness batch (M1) were not treated at all. The particularity of this experiment is the usage of a homeopath treatment with the French product Metricurine;

experimentul 2 – was made on three batches of 10 cows each, diagnosed with luteal body persistence. The cows from batch E2.1. were treated only with Estrumate® (500 mcg/dose), one administration, cows from batch E2.2. were treated with Estrumate® two doses of 2 ml i.m. (500 mcg/dose), and the witness batch was not treated;

experimentul 3 – was made on three batches of 10 cows each, for the purpose of estrus and ovulation induction and synchronization. All batches consisted of cows over 45 days after parturition. Cows from batch E3.1. were treated by application of PRIDE vaginal spirals (1,55g progesterone). Seven days later the spirals were extracted and in the 8th day Estrumate® 2 ml i.m. (500 mc/dose) was administrated. The cows from E3.2. batch received Estrumate® 2 ml i.m.500 mcg/dose) and Receptal 2,5 ml i.m. (0,01 buserelin acetate/ dose), while the witness batch (M3) was not treated.


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7.2. RESULTS AND DISCUSSIONS

In the first experiment in batch E1.1., the parturition – fecund insemination period was 52 days long for 4 cows, 78 days in three cows and 106 days for one cow.

From the total animals in batch E1.1 (10 cows) eight (80%)were diagnosed as pregnant after 90 days from the artificial insemination.

In the first experiment in batch E1.2., the parturition – fecund insemination period was 59 days long for 3 cows, 80 days in two cows and 110 days for two cows.

After the comparative analysis of the three batches, was shown that batch E1.1., constituted from cows which received hormonal treatments, had the largest number of pregnant cows (80%) compared to batch E1.2. and M1 that had 60% each.

In the second experiment - observing the results in batch E.2.1., we established that the parturition – fecund insemination interval was 53 days in three cows, 74 days in two cows and 98 days in other two cows. In batch E2.2. the parturition – fecund insemination interval was 65 days for two cows, 81 days for one cow and 96 days in three cows. Comparing the three batches we established that batch E2.2. had a higher rate of pregnancy (80%). Batch E.2.1. only 60% and batch M2 70%.

In the second experiment - observing the results in batch E.3.1., we established that the parturition – fecund insemination interval was 60 days in three cows, 76 days in two cows and 96 days in other three cows. In batch E3.2. parturition – fecund insemination interval was 61 days for two cows, 86 days for one cow and 117 days in three cows. Comparing the three batches we established that batch E.3.1. had a higher rate of pregnancy (80%). Batch E.3.2. only 60% and batch M2 40%.

CHAPTER VIII GENERAL CONCLUSIONS AND RECOMANDATIONS

In farm A the fecundity index (IF) is higher than in farm B because of the

insemination index (II) was smaller and therefore, the service period was also smaller, determining a higher economicity of the farm;

In farm B, the fecundity index was smaller than in farm A, the insemination index and the service period were higher. Farm ecnomicity level was lower and the profite smaller;

The veterinarian role is indisputable in a farm, the sanitary-veterinary state of the animals being really important for the evolution of reproductive and productive life;

A farm can be considered profitable when the fecundity index is 60-70% meaning that 60-70% of the cows remained pregnant after the first insemination, when the insemination index is 1,5-2 artificial insemination/gestation, and the service period is 75-90 days. None of the farms is considered to be in these standards;

Cow breeders have to understand and respect the farm management, especially regarding the fragile equilibrium between health, nutrition and reproduction;


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The puerperium duration has direct effects over the service-period and therefore over the first fecund insemination, zootechnical indexes that express milking cow’s exploitation economicity;

The incidence analysis of puerperal diseases shows that fetal annexes retention registered a value of 10,2%, the same as uterine atonia, genital infections 19,3% and ovariopaties 20,3%;

For the cows with puerperal metropaties the pathology wasn’t always expressed with general signs, they depending of the inflammatory process gravity, the main sign being the muco-purulent, intermittent mucus expressed on the inferior angle of the vulvae, the perineum region and posterior legs;

Our researches show a high incidence of puerperal diseases along with the periparturial morbidity (41,5% of the cases); from these, the most encountered in our two farms are the puerperal infections (on all genital tract segments) usually after fetal annexes retention, uterine prolapse, postpartum uterine atonia and ovarian pathology;

Regarding the influence of puerperal affections over the reproductions parameters we concluded that the medium parturition-first insemination period was 70,1 days for the witness batch, 122 days for the cows with postpartum infections and 121 days for the cows with postpartum ovariopaties. Service-period registered higher values for the females with pathologies than the witness batch. The same aspects are encountered for the calving interval;

Hematological results show the high difference of hemoglobin values between E1 and M1, explained by the high productivity and exploitation of the animals;

VEM and HEM presented maximal values for both batch E1 and M1, explained by body’s tendency of compensating the anemia by erythrocyte blood release and the high productivity;

For the cows with ovarian pathology (batch E2) the erythrocyte constants showed different value variations between M1 and the reference quote. Hemoglobin and hematocrit levels showed minimal values for batch E2, hemoglobin had the average value under the inferior limit of reference quote, indicating a variable anemia of cows;

The hematological values are situated in the bovine physiological limits, but in batches E1 and E2, with pathology, the average total serum proteins value was lower than witness batch and the reference value indicating protein metabolism disequilibrium;

The metabolic profile allows an early recognition of hematologic and biochemical deficiencies and permit taking correction actions for the multiple errors and therefore increasing the productive and reproductive rating;

Total Germ Number increase progressively in the first weeks post-partum, until maximal values, with significantly higher values for the cows with dystocia parturitions;

The isolated bacteria species were Gram+ in 48% of cases, and Gram-in 52% , for the cows with dystocic parturitions (predominating the association culture bacteria) and 97% Gram+ for the ones with eutocic parturitions;

In the 4th week after birth, bacteria were isolated only in cows with dystocic parturitions, 50% being from Arcanobacterium pyogenes;

The bacterial flora analysis shows equilibrium between the types with gram coloration affinity, G+ bacteria predominating (52%);


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Antibiogram was determined for the majority of samples, using the micro-comprimates. Germ sensitivity was obtained by measuring the inhibition diameters. The treatments using antibiogram showed better results instead of the ones without antibiogram;

Histopatological examinations revealed vulvae-vaginal lesions with crushed tissues and pseudo-membranes, abscesses and fistulae with macrophages in phagocitosis process and inactive;

The uterine lesions present in the genital puerperal pathology are frequent in all uterine segments: cervix, uterine body, uterine horns;

Endometrial lesions are macroscopically visible by congestive uterine mucosa edematiated in the initial phase (acute endometritis) and purple alternating with uncolored portions (according to purulent center) hypotonic or atonic uterus with purulent collections in the inside and uterine walls distension in the chronic phase;

Histopatologicaly, in acute endometritis, were showed microhemoragies in the submucosa with leucodiapedesis and in the chronic endometritis, limpho-granulocitary periglandulary and intraglandulary infiltrate, difuse and centered along with microabcesses in the submucosa and periglandular macrophages in active phagocitosis process;

The lesions present in puerperal uterine affections are mostly associated with different ovarian affections, ovarian histopatological examinations being necessary;

The fecundity index is insignificantly higher in farm A instead of farm B, because of the smaller values of insemination index and service-period;

Calf loses in the four researched years were provoked by cow lack of surveillance in the last days of gestation and the unqualified interventions made in dystocyes along with bad tending and alimentation;

In farm A was a loss of 10 calves (aprox. 2500 euro) and in farm B, 15 heads (aprox. 3750 euro), a considerable loss for the two farms.

For the both farms is necessary a increase of economic efficiency by monitoring with great attention the puerperal period by sanitary-veterinary activities, improvement of technological parameters, production level increasing, production qualitative valorification and costs reduction;

The zoo-economical implications of a sanitary-veterinary uncontrolled puerperium that degenerates in pathologic has major repercussions over the reproduction and production;

Bovine reproduction is harder to manage than for the other species because of the technological deviations that materialize firstly over the reproduction: fecundity and nativity;

Estrumate and Metricure treatments showed higher gestation rate (80%), than treatments with Metricurine (70%) and the witness batch (60%);

Two Estrumate administrations are more efficient than one E2.1. had 80% gestation rate and E2.2. had 60% gestation rate;

PRIDE spirals and Estrumate treatment (80%) is more efficient than the treatment with Estrumate and Receptal (60%);

Associated therapy contributed to an increase of gestation rate until 80% for Estrumate and Metricure or PRIDE spirals and Estrumate.


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Recomandations We recommend heifers introduction reproduction planification, for avoiding growth stopping, dystocic parturitions, low milk production and ovarian cysts; Screening of cows and heifers in heats with their individualization by using technological tablets and insemination and calving plans; Choosing the right insemination moment, before the heats end, cow fecundity being dependent of the insemination moment; Grouped calving for milking cows in spring time, for green mass valorification, low milk costs, small investments in the farm, a rational development of the youth; As a therapeutical conduct for puerperal affection prevention is important to respect the following basic elements: sporadic hematologic, metabolic and vitamin-mineral profile determination, conclusive for health status determination, gestations surveillance, equilibrated and differentiated ratio for the cows in the last gestation period;

summary of the phd thesis · of puerperium over the reproductional activities in two farms (farm a...

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